IGEM:IMPERIAL/2007/DNA Constructs: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
mNo edit summary
 
(40 intermediate revisions by 4 users not shown)
Line 1: Line 1:
__NOTOC__
<!-- Heading -->
{| style="width:800px; height:80px;" cellpadding="0" border="0"
|-
| width="220px" | <html><a href="http://www.imperial.ac.uk"><img src="http://www3.imperial.ac.uk/portal/pls/portallive/docs/1/281904.GIF" height="55"></a></html>
| width="390px" |  <br><center><html><h3>THE IMPERIAL iGEM 2007 Team </h3></html></center>
| width="110px"| <html><a href="http://www.imperial.ac.uk/centenary"><p align="right"><img src="http://www3.imperial.ac.uk/portal/pls/portallive/docs/1/7813700.GIF" height="60"></p></a></html>
|}
<!-- Nav bar -->
{| style="width:800px;" cellpadding="2" cellspacing="0" align="center"
|- style="background:#000000; color:white;" align="center"
| [[IGEM:IMPERIAL/2007|<font face="Arial" size=1 style="color:#ffffff"> '''Home'''</font>]]
| [[IGEM:IMPERIAL/2007/Projects/Cell By Date|<font face="Arial" size=1 style="color:#ffffff"> '''Cell By Date'''</font>]]
| [[IGEM:IMPERIAL/2007/Biofilm Detector|<font face="Arial" size=1 style="color:#ffffff"> '''Infector Detector'''</font>]]
| [[IGEM:IMPERIAL/2007/Wet Lab|<font face="Arial" size=1 style="color:#ffffff"> '''Wet Lab'''</font>]]
| [[IGEM:IMPERIAL/2007/Dry Lab|<font face="Arial" size=1 style="color:#ffffff"> '''Dry Lab'''</font>]]
| [[IGEM:IMPERIAL/2007/Resources|<font face="Arial" size=1 style="color:#ffffff">'''Resources'''</font>]]
| [[IGEM:IMPERIAL/2007/Meet The Team|<font face="Arial" size=1 style="color:#ffffff"> '''Meet the
Team'''</font>]]
| [[IGEM:IMPERIAL/2007/Entertainment|<font face="Arial" size=1 style="color:#ffffff"> '''Entertainment'''</font>]]
| [[IGEM:IMPERIAL/2007/Acknowledgments|<font face="Arial" size=1 style="color:#ffffff"> '''Acknowledgments'''</font>]]
|}
<br>
{| cellpadding="0" border="0" align="center"
|width="200px" class="white" valign="top"|
{|class="white" align="left" cellpadding="0"
|- valign="center"
| {{Click || image=Service_course.png | link=IGEM:IMPERIAL/2007/Wet Lab/Lab Notebook| width=40px | height=40px }}
| [[IGEM:IMPERIAL/2007/Wet Lab/Lab Notebook|<font face="Arial" size=2 style="color:#000099">'''Lab Notebook'''</font>]]<br /> <font size=1>Daily Record of Wet Lab</font>
|-
| <br>
|- valign="center"
| {{Click || image=Service_protocols.png | link=IGEM:IMPERIAL/2007/Wet Lab/Protocols| width=40px | height=40px }}
| [[IGEM:IMPERIAL/2007/Wet Lab/Protocols|<font face="Arial" size=2 style="color:#000099">'''Protocols'''</font>]]<br /> <font size=1>Fabrication & Testing Protocols</font>
|-
| <br>
|- valign="center"
| {{Click || image=Service_notebook.png | link=IGEM:IMPERIAL/2007/Wet Lab/DNA Constructs| width=40px | height=40px }}
| [[IGEM:IMPERIAL/2007/Wet Lab/DNA Constructs|<font face="Arial" size=2 style="color:#000099">'''DNA Constructs'''</font>]]<br /> <font size=1>Overview of DNA Constructs</font>
|-
| <br>
|- valign="center"
| {{Click || image=Scientificwriters.png | link=IGEM:IMPERIAL/2007/Wet Lab/Shopping List| width=40px | height=40px }}
| [[IGEM:IMPERIAL/2007/Wet Lab/Shopping List|<font face="Arial" size=2 style="color:#000099">'''Shopping List'''</font>]]<br /> <font size=1>Expenditures on Materials</font>
|}
<br clear="all">
|width="560px" class="white" valign="top" |
==Constructs Overview==
==Constructs Overview==
Here are several tables listing the bricks which we are cloning, constructing and using, with their relevant details.
A [[IGEM:IMPERIAL/2007/DNA_Constructs/New|new page]] is currently being developed!
===Cloning Table===
===Cloning Table===
Status of biobricks cloned and their applications:
{| style="width:95%; background:#33CCFF; border:1px solid #00CCFF" align="center"
{| style="width:95%; background:#33CCFF; border:1px solid #00CCFF" align="center"
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
|width=10px|I
|width=10px|I
|width=20px|ID
|width=10%|Part
|width=10%|Part
|width=20%|Description
|width=17%|Description
|width=10%|Status
|width=18%|Status
|width=22%|Ligation Target
|width=12%|Ligation Target
|width="--"%|Use in Constructs
|width="--"%|Use in Constructs
<!-- T7 PROMOTER -->
<!-- PROMOTERS WITH GFP -->
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || Minipreped || --- || ''Not being used anymore''
||  || 02 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || Basic construct for chassis characterization
<!-- TETRACYCLIN PROMOTER WITH GFP-->
 
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || Minipreped || --- || Basic construct for chassis characterization
||  || 11 || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || pT7 GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF><small>''Maxi''</small></font>,  <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || CBD test construct (pT7)


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || pBad-GFP || Not Cloned || --- || Test construct for CBD (pBad)
||  || 30 || --- || pT7 (strong GFP) || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font>|| --- || T7 test-construct


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] || ptet-luxR-plux-GFP || Minipreped || --- || ''Not being used anymore''
|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || 18 || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF><small>''Midi''</small></font>, <font color=#FF6633>BL21</font> || --- || Basic construct for chassis characterization


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || Minipreped || acGFP, DsRed, luxI || CBD final (pBad), ID AHL sender, Hrp test construct
||  ||  ||  ||  ||  ||  ||
<!-- PROMOTERS ONLY -->
|- style="background:#CCFFFF" align="center"
||  || 05 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || CBD final (ptet), ID AHL sender


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_I0500|I]] || [http://parts.mit.edu/registry/index.php/Part:BBa_I0500 BBa_I0500] || pBad || Minipreped || acGFP, DsRed || CBD final construct, Hrp test construct
|| || || BBa_I719??? || pT7 || ''Ready'' || DsRedEx || CBD final construct (pT7)


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || Minipreped || lacZ, Hrp amplifier || ID final construct
||  || 07 || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || ID final construct


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI || Minipreped || ptet || ID AHL sender final construct
||  || || || || || ||
 
<!-- ID SPECIFIC BRICKS -->
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0010 BBa_R0010] || lacI-pLac || Minipreped || --- || ''Not being used anymore''
||  || 21 || [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 BBa_J23039] || ptet-luxI || <font color=#CC0000>XL1</font>, <font color=#0000FF><small>''Midi''</small></font> || --- || ID sender construct


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || Minipreped || --- || ''Not being used anymore''
||  || 24 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0062 BBa_R0062] || plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font> || DsRedEx || ID construct 2


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP || Minipreped || --- || CBD test construct (T7)
||  || 25 || [http://parts.mit.edu/registry/index.php/Part:BBa_J37032 BBa_J37032] || plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF>''Midi''</font> || --- || ID construct 2


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] || RBS-AraC-noStop || Minipreped || ptet, pBad || CBD final construct (pBad)
||  || 26 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03119 BBa_S03119] || ptet-luxR || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font> || --- || ---


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || Minipreped || AraC || CBD test construct (pBad)
||  || || || || ||  ||  
 
<!-- BRICKS WITH PLASMID VECTORS -->
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI regulated promoter || Minipreped || acGFP, DsRed || CBD final construct
||  || 27 || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || Empty pSB1A3 || '''Problematic''' || --- || ---


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_J07037 BBa_J07037] || cI and GFP || Minipreped || --- || ''Not being used anymore''
||  || ||  ||  || || ||  
 
<!-- BRICKS FOR CONSTRUCTION [NOT USEFUL] -->
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || GFP || Not cloned || cI || CBD test construct (cI)
||  || 13 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || --- || ''General part construction''


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || Not cloned || CBD promoters || CBD final reporter
||  || 17 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || <font color=#CC0000>XL1</font>, <font color=#0000FF><small>''Midi''</small></font> || --- || ''Not being used anymore''


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || Not cloned || --- || Basic construct for chassis characterization
||  || 19 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || strong RBS || <font color=#CC0000>XL1</font> || --- || ''General part construction''


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || strong RBS || Not cloned || --- || Backup part
||  || 23 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF>''Midi''</font> || pT7 || ''General part construction''


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || T7 promoter || Pending || acGFP, DsRed || CBD final construct
||  || || || || ||  ||  
 
<!-- BRICKS BEING ORDERED -->
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || acGFP || Ordered || ptet, pBad, plux || Reporter system for test constructs, Hrp
||  ||  || BBa_I719??? || DsRed Express || Ordered || ptet, plux, pT7 || Reporter system for ID, Hrp


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || DsRed || Ordered || ptet, pBad, plux || Reporter system for ID, Hrp
||  ||  || BBa_I719??? || HrpR-HrpS || Ordered || ??? || Hrp regulators


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || HrpR-HrpS-pHrpL || Ordered || Promoter, acGFP || Hrp core system
||  ||  || BBa_I719??? || pHrpL || Ordered || ??? || Hrp promoter


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || HrpV|| Ordered || Promoter, DsRed || Hrp inhibitor system
||  ||  || BBa_I719??? || HrpV || Ordered || ??? || Hrp inhibitor system


|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
||  || BBa_I719??? || lacZ || Stopped || --- || ''Not being used anymore''
||  || || || || ||  ||  
 
|}
|}


===Ligation Table===
===Ligation Table===
Status of parts ligated and their applications:
{| style="width:95%; background:#00DD33; border:1px solid #00CC33" align="center"
{| style="width:95%; background:#00DD33; border:1px solid #00CC33" align="center"
|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
|width=10px|I
|width=10px|I
|width=20px|ID
|width=10%|Part
|width=10%|Part
|width=20%|Description
|width=17%|Description
|width=10%|Status
|width=18%|Status
|width=22%|Parents
|width=22%|Parents
|width="--"%|Application
|width="--"%|Application
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || RBS-araC & stop codon || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] + [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || CBD test construct 3 and final constructs 5, 8 and 10
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || Ptet & araC || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] + araC with stop || CBD test construct 3 and final construct 8
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || Ptet-araC & pBad-gfp || Pending || Ptet-araC + [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || CBD test construct 3
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || PcI & gfp || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] +  [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || CBD test construct 4
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || PcI & araC || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] + araC with stop || CBD test construct 5 and final construct 10
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || PcI-araC & pBad-gfp || Pending || PcI-araC + [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || CBD test construct 5
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || pT7 & DsRed || Pending || pT7 + DsRed || CBD final construct 6
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || ptet & DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040]+ DsRed || CBD final construct 7
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || pBad & DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_I0500 BBa_I0500] + DsRed || CBD final construct 8 and 10
|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || Ptet-araC & pBad-DsRed || Pending || Ptet-araC + pBad-DsRed || CBD final construct 8
||  ||  || BBa_I719??? || pT7 & GFP || Planned || pT7 + [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || CBD test construct (pT7)


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || pcI-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] + DsRed || CBD final construct 9
||  || || || || ||  ||


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || PcI-araC & pBad-DsRed || Pending || PcI-araC + pBad-DsRed || CBD final construct 10
||  ||  || BBa_I719??? || DsRedEx & pSB1A3 || ''Paused'' || DsRedEx + [http://parts.mit.edu/registry/index.php/Part:BBa_P1010 BBa_P1010] || Biobrick Submission


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || phrpL-GFP || Pending || pHrpL + GFP || ID AHL detector test construct
||  || || || || ||  ||


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || ptet-luxI || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 R0040] + [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 F1610] || ID AHL sender final construct
||  ||  || BBa_I719??? || pT7 & DsRedEx || Planned || pT7 + DsRedEx || CBD final construct (pT7)


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || ptet-luxR-plux-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + DsRed || ID AHL detector final construct
||  ||  || BBa_I719??? || ptet & DsRedEx || Planned || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] + DsRedEx || CBD final construct (ptet)


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || ptet-luxR-plux-hrpR-hrpS || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + HrpR-HrpS || ID AHL detector final construct
||  ||  || BBa_I719??? || ptet-luxR-plux-DsRedEx || Planned || [http://parts.mit.edu/registry/index.php/Part:BBa_T2620 BBa_F2620] + DsRedEx || ID detector final construct 1


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  || BBa_I719??? || phrpL-DsRed || Pending || pHrpL + DsRed || ID AHL detector final construct
||  ||  || BBa_I719??? || plux-DsRedEx || Planned || [http://parts.mit.edu/registry/index.php/Part:BBa_R0062 BBa_R0062] + DsRedEx || ID detector final construct 2


|- style="background:#CCFFCC" align="center"
|- style="background:#CCFFCC" align="center"
||  ||  ||  ||  ||  ||
||  ||  ||  ||  ||  ||  ||
|}
|}


<font size="-2">Legend: I (Information)</font>
<font size="-2">Legend: I (Information), [Status: <font color=#CC0000>XL1</font> - cloned in XL1-Blue; <font color=#CC00FF>''Maxi''</font> - maxiprepped; <font color=#0000FF>''Midi''</font> - midiprepped; <font color=#00CC00>''Mini''</font> - miniprepped; <font color=#FF6633>BL21</font> - cloned in BL21(DE3); <small>status</small> - stocks running low]</font>


===Parts from the Registry===
===Parts from the Registry===
Physical information of biobricks:
{| style="width:95%; background:#FF3399; border:1px solid #FF0099" align="center"
{| style="width:95%; background:#FF3399; border:1px solid #FF0099" align="center"
|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
Line 162: Line 195:
|width=10px| G
|width=10px| G
|width=10px| S
|width=10px| S
|width=20px|ID
|width=10%| Name
|width=10%| Name
|witdh="--"| Description
|witdh="--"| Description
|width=10%| Plasmid
|width=9%| Plasmid
|width=55px| Resist
|width=55px| Resist
|width=55px| Copy
|width=55px| Copy
Line 170: Line 204:
|width=65px| Size(T)
|width=65px| Size(T)
|width=12%| Location
|width=12%| Location
|width=7%| Conc. <br> (ng/ul)
|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 720bp || 2799bp || Well 5H, Plate 1
|| A || <font color=#0000FF>W</font> || G ||  || 02 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || GFP under tetracyclin promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 937bp || 3016bp || Well 15H, Plate 2 || 500


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || GFP under tetracyclin promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 937bp || 3016bp || Well 15H, Plate 2
|| A || W || G ||  || 05 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || tetracyclin promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 54bp || 2133bp || Well 7O, Plate 1 || 60


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || ||  || || [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || GFP under arabinose promoter || [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] || KanR || Low || 2093bp || 6518bp || Well 6H, Plate 3
|| A || W || G ||  || 07 || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || lux promoter, with luxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 1061bp || 3140bp || Well 6B, Plate 1 || 80


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] || GFP under lux promoter, with luxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 1787bp || 3866bp || Well 18K, Plate 3
|| A || <font color=#990033>X</font> || <font color=#339900>G</font> ||  || 11 || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP|| [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 826bp || 2905bp || Well 13F, Plate 2  || 100
<!-- Number 05 -->
|- style="background:#FFCCFF" align="center"
|| A || W || <font color=#990033>X</font> ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || tetracyclin promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 54bp || 2133bp || Well 7O, Plate 1


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I0500 BBa_I0500] || arabinose promoter || [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] || KanR || Low || 1210bp || 5635bp || Well 9I, Plate 2
|| A || W || G ||  || 13 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || [http://parts.mit.edu/registry/index.php/Part:pSB1AK3 pSB1AK3] || A+KR || High || 129bp || 3316bp || Well 1B, Plate 2 || 100/70


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || lux promoter, with luxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 1061bp || 3140bp || Well 6B, Plate 1
|| A || || G ||  || 17 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 861bp || 2940bp || Well 16N, Plate 1 || 80


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI coding region || [http://parts.mit.edu/registry/index.php/Part:pSB1AK3 pSB1AK3] || A+KR || High || 798bp || 3987bp || Well 1B, Plate 2
|| A || <font color=#0000FF>W</font> || G ||  || 18 || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || GFP under lux promoter with LuxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || AmpR || High || 1945bp || 4102bp || Well 19B, Plate 2 || 460


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0010 BBa_R0010] || lac promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 200bp || 2279bp || Well 7K, Plate 1
|| A || W || ||  || 19 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || RBS || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 12bp || 2091bp || Well 3O, Plate 1 || --
<!-- Number 10 -->
|- style="background:#FFCCFF" align="center"
|| A ||  || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] || KanR ||  Low || 681bp || 5106bp || Well 15M, Plate 2


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP|| [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 826bp || 2905bp || Well 13F, Plate 2
|| A || || <font color=#339900>G</font> ||  || 21 || [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 BBa_J23039] || Constitutive luxI || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 860bp || 2939bp || Well 11K, Plate 3 || 140


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || || <font color=#339900>G</font> ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] || AraC with RBS but no stop || [http://parts.mit.edu/registry/index.php/Part:pSB1AC3 pSB1AC3] || A+CR || High || 958bp || 4013bp || Well 20E, Plate 3
|| A || W || G ||  || 23 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || GFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 875bp || 2954bp || Well 16E, Plate 1 || 120/330


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || G ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || [http://parts.mit.edu/registry/index.php/Part:pSB1AK3 pSB1AK3] || A+KR || High || 129bp || 3316bp || Well 1B, Plate 2
|| A || || G ||  || 24 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0062 BBa_R0062] || lux promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 55bp || 2134bp || Well 9G, Plate 1 || 90


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W || <font color=#990033>X</font> ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 49bp || 2128bp || Well 9C, Plate 1
|| A || || G ||  || 25 || [http://parts.mit.edu/registry/index.php/Part:BBa_J37032 BBa_J37032] || GFP under lux promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 938bp || 3017bp || Well 8D, Plate 3 || 125/330


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A ||  || <font color=#339900>G</font> ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_J07037 BBa_J07037] || GFP under cI promoter || [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] || KanR || Low || 912bp || 5337bp || Well 12D, Plate 2
|| A ||  || G ||  || 26 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03119 BBa_S03119] || constitutive LuxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 998bp || 3077bp || Well 19J, Plate 1 || 60


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || ||  || || [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || GFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 876bp || 2955bp || Well 16A, Plate 1
|| A || W || <font color=#990033>X</font> ||  || 27 || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || pSB1A3 (blank) || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || Amp || High || 0bp || 2157bp || Well 23E, Plate 1 || --


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A ||  ||  || || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 861bp || 2940bp || Well 16N, Plate 1
|| || <font color=#00CC00>W</font> ||  ||  || 30 || --- || T7 with GFP(strong) || --- || Kan? || ??? || --- || ??? || --- || 150


|- style="background:#FFCCFF" align="center"
|- style="background:#FFCCFF" align="center"
|| A || W ||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || GFP under lux promoter, with luxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || AmpR || High ||  1945bp || 4102bp || Well 19B, Plate 2
||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||
 
|- style="background:#FFCCFF" align="center"
|| A || W ||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || RBS || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 12bp || 2091bp || Well 3O, Plate 1
 
|- style="background:#FFCCFF" align="center"
||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  
|}
|}
<font size="-2">Legend: A (Available), W (Working), G (Checked on gel) [<font color=#339900>G</font> - incorrect size; <font color=#990033>X</font> - does not appear on gel], S (Sequenced)</font>


===Our Parts===
===Our Parts===
Physical information of our ligated parts:
{| style="width:95%; background:#FF9933; border:1px solid #FF6633" align="center"
{| style="width:95%; background:#FF9933; border:1px solid #FF6633" align="center"
|- style="background:#FFFFBB" align="center"
|- style="background:#FFFFBB" align="center"
Line 239: Line 262:
|width=10px| G
|width=10px| G
|width=10px| S
|width=10px| S
|width=20px| ID
|width=10%| Name
|width=10%| Name
|witdh="--"| Description
|witdh="--"| Description
Line 248: Line 272:
|width=12%| Stock
|width=12%| Stock
|- style="background:#FFFFBB" align="center"
|- style="background:#FFFFBB" align="center"
||  ||  ||  ||  || BBa_I791??? || luxI coding region under tetracyclin promoter || pSB1A2 || AmpR || High || 860bp || 2939bp ||
||  ||  ||  ||  || || [http://parts.mit.edu/registry/index.php/Part:BBa_I719022 BBa_I719022] || cI with GFP || pSB1A2 || AmpR || High || 933bp || 3988bp ||


|- style="background:#FFFFBB" align="center"
|- style="background:#FFFFBB" align="center"
||  ||  ||  ||  || BBa_I791??? || DsRed under tetracyclin promoter || pSB1A2 || AmpR || High || || ||
||  ||  ||  ||  || || BBa_I791021 || luxI coding region under ptet || pSB1A2 || AmpR || High || 860bp || 2939bp ||


|- style="background:#FFFFBB" align="center"
|- style="background:#FFFFBB" align="center"
||  ||  ||  ||  || BBa_I791??? || DsRed under arabinose promoter || pSB2K3 || KanR || Low ||  ||  ||
||  ||  ||  ||  ||  || BBa_I791??? || DsRed Express under ptet || pSB1A2 || AmpR || High ||  ||  ||


|- style="background:#FFFFBB" align="center"
|- style="background:#FFFFBB" align="center"
||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||
||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||  ||
|}
|}
<font size="-2">Legend: A (Available), W (Working) [<font color=#00CC00>W</font> - working ''in vivo'', <font color=##0000FF>W</font> - working ''in vitro'', <font color=#990033>X</font> - faulty], G (Checked on gel) [<font color=#339900>G</font> - incorrect size; <font color=#990033>X</font> - does not appear on gel], S (Sequenced)<br>Concentrations are given in ng/μl</font>


===Part Graveyard===
===Part Graveyard===
General information on parts we do not like...
{| style="width:95%; background:#666666; border:1px solid #444444" align="center"
{| style="width:95%; background:#666666; border:1px solid #444444" align="center"
|- style="background:#DDDDDD" align="center"
|- style="background:#DDDDDD" align="center"
|width=10px| A
|width=10px| A
|width=10px| W
|width=10px| W
|width=20px| ID
|width=10%| Name
|width=10%| Name
|witdh=30%| Description
|witdh=30%| Description
|width=10%| Plasmid
|width=10%| Plasmid
|width=65px| Size
|width=65px| Size
|width=10%| Status
|width=15%| Status
|width="--"%| Notes
|width="--"%| Notes
|- style="background:#DDDDDD" align="center"
|- style="background:#DDDDDD" align="center"
|| A ||  || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || Miniprepped || No RBS present on this part
|| A || W || 01 || BBa_E0040 || GFP || pSB1A2|| 720bp || XL1, Mini || Redundant
 
|- style="background:#DDDDDD" align="center"
|| A ||  || 03 || BBa_J5528 || pBad-GFP || pSB2K3 || 2093bp || failed-XL1 || ara construct abandoned
 
|- style="background:#DDDDDD" align="center"
|| A ||  || 04 || BBa_J24677 || plux-GFP || pSB1A2 || 1787bp || XL1, Mini || Wrong GFP ligated
 
|- style="background:#DDDDDD" align="center"
|| A ||  || 06 || BBa_I0500 || pBad || pSB2K3 || 1210bp || XL1, Mini, Midi || ara construct abandoned
 
|- style="background:#DDDDDD" align="center"
|| A || W || 08 || BBa_F1610 || luxI || pSB1AK3 || 798bp || XL1, Mini, Midi|| Substitute part J23039
 
|- style="background:#DDDDDD" align="center"
|| A || W || 09 || BBa_R0010  || lacI-plac || pSB1A2 || 200bp || XL1, Mini || Lac construct abandoned


|- style="background:#DDDDDD" align="center"
|- style="background:#DDDDDD" align="center"
||  ||  || || || || || ||  
|| A || W || 10 || BBa_E1010 || RFP || pSB2K3 || 681bp || XL1, Mini || Substitute part I13507
|}
 
|- style="background:#DDDDDD" align="center"
|| A ||  || 12 || BBa_S03550 || araC || pSB1AC3 || 958bp || XL1, Mini, Midi || araC construct abandoned
 
|- style="background:#DDDDDD" align="center"
||  || W || 14 || BBa_R0051 || pcI || pSB1A2 || 49bp || Biobrick || pcI construct abandoned


==Constructs for Cell By Date==
|- style="background:#DDDDDD" align="center"
|| A ||  || 15 || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || XL1, Mini || No RBS present on this part


*Construct CBD 1
|- style="background:#DDDDDD" align="center"
**Entire construct is currently available in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
|| A || W || 16 || BBa_E0240 || GFP || pSB1A2 || 876bp || XL1, Midi || Redundant
**pT7 can be used with T7 RNAP in vitro and in veso, and it gives constitutive expression.
[[image:ICGEMS_dvc_PT7_gfp.png|left|thumb|200px|PT7 promoting GFP]]<br clear="all">


*Construct CBD 2
|- style="background:#DDDDDD" align="center"
**Entire construct is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
|| || W || 20 || --- || AraC || pSB1AC3 || 1095bp || Planned || araC construct abandoned
**pTet can be used with E.coli RNAP in vitro and maybe in veso, and it gives constitutive expression.
[[image:ICGEMS_dvc_Ptet_gfp.png|left|thumb|200px|Ptet promoting GFP]]<br clear="all">


*Construct CBD 3
|- style="background:#DDDDDD" align="center"
**pBAD promoter with GFP is currently available in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] (KanR)
|| A || W || 28 || BBa_P1010 || pSB1A3 (ccd) || pSB1A3 || 0bp || failed-XL1 || Strain not available
**Need Ara C regulator protein in vitro and in veso. Ara C available with the RBS:[http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550]
**pBad can be used with E.coli RNAP in vitro, and it is inducible by arabinose.
**To be used in veso, arabinose transport proteins have to be inserted in the phospholipid bilayer of the vesicles.
[[image:ICGEMS_dvc_PBad_gfp.png|left|thumb|400px|PBAD promoting GFP and Ptet promoting araC]]<br clear="all">


*Construct CBD 4
|- style="background:#DDDDDD" align="center"
**Entire construct is currently available in Biobricks Registry:  [http://parts.mit.edu/registry/index.php/Part:BBa_J07037 BBa_J07037] (avaiable), in [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] (KanR)
|| A || W || 29 || BBa_P1010 || pSB1A2 (ccd) || pSB1A2 || 0bp || --- || Strain not available
**pcI gives constitutive expression.
[[image:PcI_construct.PNG|left|thumb|400px|PcI promoting GFP]]<br clear="all">


*Construct CBD 5
|- style="background:#DDDDDD" align="center"
**All parts are currently available in the Biobricks Registry.
|| ||  ||  ||  ||  ||  ||  ||  ||
**pcI promoter: [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
|}
**Arabinose C regulator protein with RBS: [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1AC3 pSB1AC3] (A+CR)
**Stop codon: [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015], in [http://parts.mit.edu/registry/index.php/Part:pSB1AK3 pSB1AK3] (A+KR)
**pBAD with GFP: [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528], in [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] (KanR)
[[image:ICGEMS_dvc_PBad2_gfp.PNG‎ |left|thumb|400px|PBAD promoting GFP and PcI promoting araC]] <br clear="all">


*Construct CBD 6
**Parts avaiable ....
[[Image:ICGEMS_dvc_PT7_rfp.PNG|left|thumb|200px|pT7 promoting RFP]] <br clear="all">


*Construct CBD 7
==Construct Diagrams==
**Parts avaiable ....
The original (old) diagrams page was moved to [[IGEM:IMPERIAL/2007/DNA_Constructs/Con_Diagrams_old|old construct diagrams]] page.  This page also contains the obsolete parts.
[[image:ICGEMS_dvc_Ptet_rfp.PNG|left|thumb|200px|Ptet promoting RFP]]


===CBD Constructs===
'''ptet GFP'''
[[image:ICGEMS_dvc_Ptet_gfp.png|left|thumb|200px|Ptet promoting GFP]]
*Availability: Found in registry as [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522]
<br clear="all">
<br clear="all">
*Construct CBD 8
**Parts avaiable ....
[[image:ICGEMS_dvc_PBad_rfp.PNG‎|left|thumb|400px|PBAD promoting RFP and Ptet promoting araC]]


'''pT7 GFP'''
[[image:ICGEMS_dvc_PT7_gfp.png|left|thumb|200px|PT7 promoting GFP]]
*Availability: Found in resigry as [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104]
<br clear="all">
<br clear="all">
*Construct CBD 9
**Parts avaiable ....
[[image:ICGEMS_dvc_PcI_rfp.PNG|left|thumb|200px|pcI promoting RFP]]


'''ptet DsRed express'''
[[image:ICGEMS_dvc_Ptet_rfp.PNG|left|thumb|200px|Ptet promoting dsRed]]
*Availability: Requires ligation
*Promoter can be found in registry as: [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040]
*DsRed express is to be ordered from GeneArt
<br clear="all">
<br clear="all">
*Construct CBD 10
**Parts avaiable ....
[[Image:ICGEMS_dvc_PBad2_rfp.PNG‎|left|thumb|400px|PBAD promoting RFP and PcI promoting araC]]


'''pT7 DsRed express'''
[[Image:ICGEMS_dvc_PT7_rfp.PNG|left|thumb|200px|pT7 promoting dsRed]]
*Availability: Requires ligation
*Promoter is being PCRed from plasmid derived externally
*DsRed express is to be ordered from GeneArt
<br clear="all">
<br clear="all">


==Constructs for Infector Detector==
===ID Constructs===
'''pLux GFP with LuxR'''
[[image:ICGEMS_sys_Ptet_luxR_Plux_gfp.png|left|thumb|400px|Ptet promoting LuxR, Plux promoting GFP]]
*Availability: Found in registry as [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002]
<br clear="all">


*Construct ID 1
'''pLux GFP with LuxR'''
**Entire part minus promoter is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1AK3 pSB1AK3] (AmpR, KanR)
[[image:ICGEMS_sys_Plux_gfp.png‎|left|thumb|200px|Plux promoting GFP]]
**pTet is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
*Availability: Found in registry as [http://parts.mit.edu/registry/index.php/Part:BBa_J37032 BBa_J37032]  
**pTet can be used with E.coli RNAP in vitro and maybe in veso, and it gives constitutive expression.
<br clear="all">
[[image:ICGEMS_dvc_Ptet_luxI.png|left|thumb|200px|Ptet promoting LuxI]]<br clear="all">


*Construct ID 2
''<<Two more constructs with DsRed express yet to be included>>''
**Entire part is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
**Entire part minus GFP reporter system is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] (available), in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
**GFP reporter system is currently in Biobricks Registry: BBa_I13504 (available)
**pLux can be used with E.coli RNAP in vitro and maybe in veso, and it is repressed by LuxR but activated by AHL:LuxR complex.
[[image:ICGEMS_sys_Ptet_luxR_Plux_gfp.png|left|thumb|400px|Ptet promoting LuxR, Plux promoting GFP]]<br clear="all">


*Construct ID 3
|}
**Entire part minus Hrp system is currently in Biobricks Registry: [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620](available) in [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] (AmpR)
**pHrpL can be used with E.coli RNAP in vitro and maybe in veso, and it is inducible by HrpR:HrpS:Sigma 54 complex.
[[Image:ICGEMS_sys_Ptet_luxR_Plux_hrpR-hrpS-gfp.PNG‎ |left|thumb|600px|Ptet promoting LuxR, Plux promoting HrpR and HrpS and PHrpL promoting GFP]]<br clear="all">
 
*Construct ID 4
** Parts available ...
[[Image:ICGEMS_sys_Ptet_luxR_Plux_lacz.PNG‎|left|thumb|400px|Ptet promoting LuxI]]<br clear="all">
 
*Construct ID 5
**Parts available ...
[[Image:ICGEMS_sys_Ptet_luxR_Plux_hrpR-hrpS-lacZ.PNG‎ ‎|left|thumb|600px|Ptet promoting LuxR, Plux promoting HrpR and HrpS and PHrpL promoting lacZ]]<br clear="all">

Latest revision as of 03:21, 2 October 2007

<html><a href="http://www.imperial.ac.uk"><img src="http://www3.imperial.ac.uk/portal/pls/portallive/docs/1/281904.GIF" height="55"></a></html>
<html><h3>THE IMPERIAL iGEM 2007 Team </h3></html>
 <html><a href="http://www.imperial.ac.uk/centenary"><p align="right"><img src="http://www3.imperial.ac.uk/portal/pls/portallive/docs/1/7813700.GIF" height="60"></p></a></html>
Home Cell By Date Infector Detector Wet Lab Dry Lab Resources Meet the Team Entertainment Acknowledgments


Lab Notebook
Daily Record of Wet Lab

Protocols
Fabrication & Testing Protocols

DNA Constructs
Overview of DNA Constructs

Shopping List
Expenditures on Materials


Constructs Overview

Here are several tables listing the bricks which we are cloning, constructing and using, with their relevant details.

A new page is currently being developed!

Cloning Table

Status of biobricks cloned and their applications:

I ID Part Description Status Ligation Target Use in Constructs
02 BBa_I13522 ptet-GFP XL1, Maxi, Mini, BL21 --- Basic construct for chassis characterization
11 BBa_E7104 pT7 GFP XL1, Maxi, Midi, Mini, BL21 --- CBD test construct (pT7)
30 --- pT7 (strong GFP) XL1, Maxi, Mini, BL21 --- T7 test-construct
I 18 BBa_T9002 ptet-luxR-plux-GFP XL1, Maxi, Midi, BL21 --- Basic construct for chassis characterization
05 BBa_R0040 ptet XL1, Midi, Mini DsRedEx CBD final (ptet), ID AHL sender
BBa_I719??? pT7 Ready DsRedEx CBD final construct (pT7)
07 BBa_F2620 ptet-luxR-plux XL1, Midi, Mini DsRedEx ID final construct
21 BBa_J23039 ptet-luxI XL1, Midi --- ID sender construct
24 BBa_R0062 plux XL1, Midi DsRedEx ID construct 2
25 BBa_J37032 plux-GFP XL1, Maxi, Midi --- ID construct 2
26 BBa_S03119 ptet-luxR XL1, Midi --- ---
27 pSB1A3 Empty pSB1A3 Problematic --- ---
13 BBa_B0015 Double terminator XL1, Maxi, Midi, Mini --- General part construction
17 BBa_I13507 RFP XL1, Midi --- Not being used anymore
19 BBa_B0034 strong RBS XL1 --- General part construction
23 BBa_I13504 GFP XL1, Maxi, Midi pT7 General part construction
BBa_I719??? DsRed Express Ordered ptet, plux, pT7 Reporter system for ID, Hrp
BBa_I719??? HrpR-HrpS Ordered ??? Hrp regulators
BBa_I719??? pHrpL Ordered ??? Hrp promoter
BBa_I719??? HrpV Ordered ??? Hrp inhibitor system

Ligation Table

Status of parts ligated and their applications:

I ID Part Description Status Parents Application
BBa_I719??? pT7 & GFP Planned pT7 + BBa_I13504 CBD test construct (pT7)
BBa_I719??? DsRedEx & pSB1A3 Paused DsRedEx + BBa_P1010 Biobrick Submission
BBa_I719??? pT7 & DsRedEx Planned pT7 + DsRedEx CBD final construct (pT7)
BBa_I719??? ptet & DsRedEx Planned BBa_R0040 + DsRedEx CBD final construct (ptet)
BBa_I719??? ptet-luxR-plux-DsRedEx Planned BBa_F2620 + DsRedEx ID detector final construct 1
BBa_I719??? plux-DsRedEx Planned BBa_R0062 + DsRedEx ID detector final construct 2

Legend: I (Information), [Status: XL1 - cloned in XL1-Blue; Maxi - maxiprepped; Midi - midiprepped; Mini - miniprepped; BL21 - cloned in BL21(DE3); status - stocks running low]

Parts from the Registry

Physical information of biobricks:

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(T) Location Conc.
(ng/ul)
A W G 02 BBa_I13522 GFP under tetracyclin promoter pSB1A2 AmpR High 937bp 3016bp Well 15H, Plate 2 500
A W G 05 BBa_R0040 tetracyclin promoter pSB1A2 AmpR High 54bp 2133bp Well 7O, Plate 1 60
A W G 07 BBa_F2620 lux promoter, with luxR pSB1A2 AmpR High 1061bp 3140bp Well 6B, Plate 1 80
A X G 11 BBa_E7104 T7 GFP pSB1A2 AmpR High 826bp 2905bp Well 13F, Plate 2 100
A W G 13 BBa_B0015 Double terminator pSB1AK3 A+KR High 129bp 3316bp Well 1B, Plate 2 100/70
A G 17 BBa_I13507 RFP pSB1A2 AmpR High 861bp 2940bp Well 16N, Plate 1 80
A W G 18 BBa_T9002 GFP under lux promoter with LuxR pSB1A3 AmpR High 1945bp 4102bp Well 19B, Plate 2 460
A W 19 BBa_B0034 RBS pSB1A2 AmpR High 12bp 2091bp Well 3O, Plate 1 --
A G 21 BBa_J23039 Constitutive luxI pSB1A2 AmpR High 860bp 2939bp Well 11K, Plate 3 140
A W G 23 BBa_I13504 GFP pSB1A2 AmpR High 875bp 2954bp Well 16E, Plate 1 120/330
A G 24 BBa_R0062 lux promoter pSB1A2 AmpR High 55bp 2134bp Well 9G, Plate 1 90
A G 25 BBa_J37032 GFP under lux promoter pSB1A2 AmpR High 938bp 3017bp Well 8D, Plate 3 125/330
A G 26 BBa_S03119 constitutive LuxR pSB1A2 AmpR High 998bp 3077bp Well 19J, Plate 1 60
A W X 27 pSB1A3 pSB1A3 (blank) pSB1A3 Amp High 0bp 2157bp Well 23E, Plate 1 --
W 30 --- T7 with GFP(strong) --- Kan? ??? --- ??? --- 150

Our Parts

Physical information of our ligated parts:

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(P) Stock
BBa_I719022 cI with GFP pSB1A2 AmpR High 933bp 3988bp
BBa_I791021 luxI coding region under ptet pSB1A2 AmpR High 860bp 2939bp
BBa_I791??? DsRed Express under ptet pSB1A2 AmpR High

Legend: A (Available), W (Working) [W - working in vivo, W - working in vitro, X - faulty], G (Checked on gel) [G - incorrect size; X - does not appear on gel], S (Sequenced)
Concentrations are given in ng/μl

Part Graveyard

General information on parts we do not like...

A W ID Name Description Plasmid Size Status Notes
A W 01 BBa_E0040 GFP pSB1A2 720bp XL1, Mini Redundant
A 03 BBa_J5528 pBad-GFP pSB2K3 2093bp failed-XL1 ara construct abandoned
A 04 BBa_J24677 plux-GFP pSB1A2 1787bp XL1, Mini Wrong GFP ligated
A 06 BBa_I0500 pBad pSB2K3 1210bp XL1, Mini, Midi ara construct abandoned
A W 08 BBa_F1610 luxI pSB1AK3 798bp XL1, Mini, Midi Substitute part J23039
A W 09 BBa_R0010 lacI-plac pSB1A2 200bp XL1, Mini Lac construct abandoned
A W 10 BBa_E1010 RFP pSB2K3 681bp XL1, Mini Substitute part I13507
A 12 BBa_S03550 araC pSB1AC3 958bp XL1, Mini, Midi araC construct abandoned
W 14 BBa_R0051 pcI pSB1A2 49bp Biobrick pcI construct abandoned
A 15 BBa_J07037 cI-GFP pSB2K3 912bp XL1, Mini No RBS present on this part
A W 16 BBa_E0240 GFP pSB1A2 876bp XL1, Midi Redundant
W 20 --- AraC pSB1AC3 1095bp Planned araC construct abandoned
A W 28 BBa_P1010 pSB1A3 (ccd) pSB1A3 0bp failed-XL1 Strain not available
A W 29 BBa_P1010 pSB1A2 (ccd) pSB1A2 0bp --- Strain not available


Construct Diagrams

The original (old) diagrams page was moved to old construct diagrams page. This page also contains the obsolete parts.

CBD Constructs

ptet GFP

Ptet promoting GFP


pT7 GFP

PT7 promoting GFP


ptet DsRed express

Ptet promoting dsRed
  • Availability: Requires ligation
  • Promoter can be found in registry as: BBa_R0040
  • DsRed express is to be ordered from GeneArt


pT7 DsRed express

pT7 promoting dsRed
  • Availability: Requires ligation
  • Promoter is being PCRed from plasmid derived externally
  • DsRed express is to be ordered from GeneArt


ID Constructs

pLux GFP with LuxR

Ptet promoting LuxR, Plux promoting GFP


pLux GFP with LuxR

Plux promoting GFP


<<Two more constructs with DsRed express yet to be included>>

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:IMPERIAL/2007/DNA_Constructs&oldid=155665"