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

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||  || 01 || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || <font color=#CC0000>XL1</font>, <font color=#00CC00>''Mini''</font> || --- || ''Not being used anymore''
||  || 01 || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || <font color=#CC0000>XL1</font>, <font color=#00CC00>''Mini''</font> || --- || ''Not being used anymore''
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||  || 10 || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || <font color=#CC0000>XL1</font>, <font color=#00CC00>''Mini''</font> || --- || ''Not being used anymore''


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|| A || W || G ||  || 08 || [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 || W || G ||  || 08 || [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
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|| A ||  || G ||  || 10 || [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


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|width=10%| Plasmid
|width=10%| Plasmid
|width=65px| Size
|width=65px| Size
|width=10%| Status
|width=15%| Status
|width="--"%| Notes
|width="--"%| Notes
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|| A ||  || 03 || BBa_J5528 || pBad-GFP || pSB2K3 || 2093bp || Failed || ara construct abandoned
|| A ||  || 03 || BBa_J5528 || pBad-GFP || pSB2K3 || 2093bp || failed-XL1 || ara construct abandoned
 
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|| A ||  || 04 || BBa_J24677 || plux-GFP || pSB1A2 || 1787bp || XL1, Mini || Wrong GFP ligated


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|| A ||  || 04 || BBa_J24677 || plux-GFP || pSB1A2 || 1787bp || Miniprepped || Wrong GFP ligated
|| A ||  || 06 || BBa_I0500 || pBad || pSB2K3 || 1210bp || XL1, Mini, Midi || ara construct abandoned


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|| A || || 06 || BBa_I0500 || pBad || pSB2K3 || 1210bp || Midiprepped|| ara construct abandoned
|| A || W || 08 || BBa_F1610 || luxI || pSB1AK3 || 798bp || XL1, Mini, Midi|| Substitute part J23039


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|| A || W || 08 || BBa_F1610 || luxI || pSB1AK3 || 798bp || Midiprepped || Substitute part J23039
|| A || W || 09 || BBa_R0010  || lacI-plac || pSB1A2 || 200bp || XL1, Mini || Lac construct abandoned


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|| A || W || 09 || BBa_R0010  || lacI-plac || pSB1A2 || 200bp || Miniprepped || Lac construct abandoned
|| A || W || 10 || BBa_E1010 || RFP || pSB2K3 || 681bp || XL1, Mini ||


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|| A ||  || 12 || BBa_S03550 || araC || pSB1AC3 || 958bp || Midiprepped || araC construct abandoned
|| A ||  || 12 || BBa_S03550 || araC || pSB1AC3 || 958bp || XL1, Mini, Midi || araC construct abandoned


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|| A ||  || 15 || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || Miniprepped || No RBS present on this part
|| A ||  || 15 || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || XL1, Mini || No RBS present on this part


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|| A || W || 16 || BBa_E0240 || GFP || pSB1A2 || 876bp || Midiprepped || Redundant
|| A || W || 16 || BBa_E0240 || GFP || pSB1A2 || 876bp || XL1, Midi || Redundant


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|- style="background:#DDDDDD" align="center"

Revision as of 11:23, 15 August 2007

Constructs Overview

Cloning Table

I ID Part Description Status Ligation Target Use in Constructs
02 BBa_I13522 ptet-GFP XL1, Midi, Mini, BL21 --- Basic construct for chassis characterization
11 BBa_E7104 T7 GFP XL1, Midi, Mini, BL21 --- CBD test construct (T7)
I 18 BBa_T9002 ptet-luxR-plux-GFP XL1, Midi --- Basic construct for chassis characterization
22 BBa_S03335 cI-GFP Not Cloned --- CBD test construct (cI)
05 BBa_R0040 ptet XL1, Midi, Mini DsRed CBD final (pBad), ID AHL sender, Hrp test construct
BBa_I719??? T7 promoter Pending DsRed CBD final construct
07 BBa_F2620 ptet-luxR-plux XL1, Midi, Mini DsRed ID final construct
14 BBa_R0051 λcI regulated promoter XL1, Midi, Mini DsRed CBD final construct
08 BBa_F1610 luxI XL1, Midi, Mini ptet ID AHL sender final construct
21 BBa_J23039 ptet-LuxI Not Cloned --- ID sender construct
01 BBa_E0040 GFP XL1, Mini --- Not being used anymore
13 BBa_B0015 Double terminator XL1, Midi, Mini --- General part construction
17 BBa_I13507 RFP XL1, Midi --- Not being used anymore
19 BBa_B0034 strong RBS XL1, Waiting Midi --- General part construction
BBa_I719??? acGFP Ordered ptet, plux, cI Reporter system for test constructs, Hrp
BBa_I719??? DsRed Ordered ptet, plux, cI Reporter system for ID, Hrp
BBa_I719??? HrpR-HrpS-pHrpL Ordered ??? Hrp core system
BBa_I719??? HrpV Ordered ??? Hrp inhibitor system

Ligation Table

I ID Part Description Status Parents Application
BBa_I719021 ptet & luxI Concerns R0040 + F1610 ID final sender (duplicated - J23039)
BBa_I719022 cI & GFP Concerns BBa_R0051 + BBa_E0240 CBD test construct (cI)
BBa_I719??? pT7 & DsRed Pending pT7 + DsRed CBD final construct (pT7)
BBa_I719??? ptet & DsRed Pending BBa_R0040+ DsRed CBD final construct (ptet)
BBa_I719??? pcI-DsRed Pending BBa_R0051 + DsRed CBD final construct (pcI)
BBa_I719??? ptet-luxR-plux-DsRed Pending BBa_F2620 + DsRed ID detector final construct
BBa_I719??? hrpR-hrpS & phrpL Pending hrpR-hrpS + phrpL ID detector test construct 3 and final construct 5
BBa_I719??? Ptet-luxR-Plux & hrpR-hrpS-phrpL Pending BBa_F2620 + hrpR-hrpS-phrpL ID detector test construct 3 and final construct 5
BBa_I719??? Ptet-luxR-Plux-hrpR-hrpS-phrpL & GFP Pending Ptet-luxR-Plux-hrpR-hrpS-phrpL + BBa_E0240 ID detector test construct 3
BBa_I719??? Ptet-luxR-Plux-hrpR-hrpS-phrpL & DsRed Pending Ptet-luxR-Plux-hrpR-hrpS-phrpL + DsRed ID detector final construct 5


Legend: I (Information)

Parts from the Registry

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(T) Location
A W G 01 BBa_E0040 GFP pSB1A2 AmpR High 720bp 2799bp Well 5H, Plate 1
A W G 02 BBa_I13522 GFP under tetracyclin promoter pSB1A2 AmpR High 937bp 3016bp Well 15H, Plate 2
A W X 05 BBa_R0040 tetracyclin promoter pSB1A2 AmpR High 54bp 2133bp Well 7O, Plate 1
A W G 07 BBa_F2620 lux promoter, with luxR pSB1A2 AmpR High 1061bp 3140bp Well 6B, Plate 1
A W G 08 BBa_F1610 luxI coding region pSB1AK3 A+KR High 798bp 3987bp Well 1B, Plate 2
A W G 11 BBa_E7104 T7 GFP pSB1A2 AmpR High 826bp 2905bp Well 13F, Plate 2
A W G 13 BBa_B0015 Double terminator pSB1AK3 A+KR High 129bp 3316bp Well 1B, Plate 2
A W X 14 BBa_R0051 λcI promoter pSB1A2 AmpR High 49bp 2128bp Well 9C, Plate 1
A 17 BBa_I13507 RFP pSB1A2 AmpR High 861bp 2940bp Well 16N, Plate 1
A W 18 BBa_T9002 GFP under lux promoter, with luxR pSB1A3 AmpR High 1945bp 4102bp Well 19B, Plate 2
A W 19 BBa_B0034 RBS pSB1A2 AmpR High 12bp 2091bp Well 3O, Plate 1
A 21 BBa_J23039 Constitutive luxI pSB1A2 AmpR High 860bp 2939bp Well 11K, Plate 3

Legend: A (Available), W (Working), G (Checked on gel) [G - incorrect size; X - does not appear on gel], S (Sequenced)

Our 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 tetracyclin promoter pSB1A2 AmpR High 860bp 2939bp
BBa_I791??? DsRed under tetracyclin promoter pSB1A2 AmpR High
BBa_I791??? DsRed under arabinose promoter pSB2K3 KanR Low

Part Graveyard

A W ID Name Description Plasmid Size Status Notes
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
A 12 BBa_S03550 araC pSB1AC3 958bp XL1, Mini, Midi araC 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
20 BBa_I791020 AraC pSB1AC3 1095bp Planned araC construct abandoned

Constructs for Cell By Date

  • Construct CBD 1
    • Entire construct is currently available in Biobricks Registry: BBa_E7104 (available), in pSB1A2 (AmpR)
    • pT7 can be used with T7 RNAP in vitro and in veso, and it gives constitutive expression.
PT7 promoting GFP


  • Construct CBD 2
    • Entire construct is currently in Biobricks Registry: BBa_I13522 (available), in pSB1A2 (AmpR)
    • pTet can be used with E.coli RNAP in vitro and maybe in veso, and it gives constitutive expression.
Ptet promoting GFP


  • Construct CBD 3
    • pBAD promoter with GFP is currently available in Biobricks Registry: BBa_J5528 (available), in pSB2K3 (KanR)
    • Need Ara C regulator protein in vitro and in veso. Ara C available with the RBS: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.
PBAD promoting GFP and Ptet promoting araC


  • Construct CBD 4
    • Entire construct is currently available in Biobricks Registry: BBa_J07037 (avaiable), in pSB2K3 (KanR)
    • pcI gives constitutive expression.
PcI promoting GFP


PBAD promoting GFP and PcI promoting araC


  • Construct CBD 6
    • T7 promoter is in the process of cloning
    • dsRed has been ordered
pT7 promoting dsRed


  • Construct CBD 7
    • tet promoter available in the registry: BBa_R0040
    • dsRed has been ordered
Ptet promoting dsRed


  • Construct CBD 8
PBAD promoting RFP and Ptet promoting araC


  • Construct CBD 9
    • λcI regulated promoter available in the registry:BBa_R0051
pcI promoting RFP


  • Construct CBD 10
    • Parts avaiable ....
PBAD promoting RFP and PcI promoting araC


Constructs for Infector Detector

  • Construct ID 1
    • Entire part minus promoter is currently in Biobricks Registry: BBa_F1610 (available), in pSB1AK3 (AmpR, KanR)
    • pTet is currently in Biobricks Registry: BBa_R0040 (available), in pSB1A2 (AmpR)
    • pTet can be used with E.coli RNAP in vitro and maybe in veso, and it gives constitutive expression.
Ptet promoting LuxI


  • Construct ID 2
    • Entire part is currently in Biobricks Registry: BBa_J24677 (available), in pSB1A2 (AmpR)
    • Entire part minus GFP reporter system is currently in Biobricks Registry: BBa_F2620 (available), in 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.
Ptet promoting LuxR, Plux promoting GFP


  • Construct ID 3
    • Entire part minus Hrp system is currently in Biobricks Registry: BBa_F2620(available) in 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.
Ptet promoting LuxR, Plux promoting HrpR and HrpS and PHrpL promoting GFP


  • Construct ID 4
    • Entire part minus dsRed reporter system is currently in Biobricks Registry: BBa_F2620 (available), in pSB1A2 (AmpR)
    • dsRed has been ordered
Ptet promoting LuxI


  • Construct ID 5
    • Entire part minus the hrp system and dsRed reporter system is currently in Biobricks Registry: BBa_F2620 (available), in pSB1A2 (AmpR)
    • dsRed has been ordered
    • hrpR-hrpS-pHrpL has been ordered
Ptet promoting LuxR, Plux promoting HrpR and HrpS and PHrpL promoting lacZ


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