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

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<!-- T7 PROMOTER -->
<!-- T7 PROMOTER -->
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||  || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || Miniprep || --- || ''Not being used anymore''
||  || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || Minipreped || --- || ''Not being used anymore''
<!-- TETRACYCLIN PROMOTER WITH GFP-->
<!-- TETRACYCLIN PROMOTER WITH GFP-->
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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || Miniprep || --- || Basic construct for chassis characterization
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || Minipreped || --- || Basic construct for chassis characterization


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|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] || ptet-luxR-plux-GFP || Miniprep || --- || ''Not being used anymore''
|| [[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''


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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || Miniprep || acGFP, dsRFP, luxI || CBD final (pBad), ID AHL sender, Hrp test construct
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || Minipreped || acGFP, dsRFP, luxI || CBD final (pBad), ID AHL sender, Hrp test construct


|- style="background:#CCFFFF" align="center"
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|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_I0500|I]] || [http://parts.mit.edu/registry/index.php/Part:BBa_I0500 BBa_I0500] || pBad || Miniprep || acGFP, dsRFP || CBD final construct, Hrp test construct
|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_I0500|I]] || [http://parts.mit.edu/registry/index.php/Part:BBa_I0500 BBa_I0500] || pBad || Minipreped || acGFP, dsRFP || CBD final construct, Hrp test construct


|- style="background:#CCFFFF" align="center"
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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || Miniprep || lacZ, Hrp amplifier || ID final construct
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || Minipreped || lacZ, Hrp amplifier || ID final construct


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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI || Miniprep || ptet || ID AHL sender final construct
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI || Minipreped || ptet || ID AHL sender final construct


|- style="background:#CCFFFF" align="center"
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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0010 BBa_R0010] || lacI-pLac || Miniprep || --- || ''Not being used anymore''
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0010 BBa_R0010] || lacI-pLac || Minipreped || --- || ''Not being used anymore''


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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || Miniprep || --- || ''Not being used anymore''
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || Minipreped || --- || ''Not being used anymore''


|- style="background:#CCFFFF" align="center"
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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP || Miniprep || --- || CBD test construct (T7)
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP || Minipreped || --- || CBD test construct (T7)


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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] || RBS-AraC-noStop || Miniprep || ptet, pBad || CBD final construct (pBad)
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_S03550 BBa_S03550] || RBS-AraC-noStop || Minipreped || ptet, pBad || CBD final construct (pBad)


|- style="background:#CCFFFF" align="center"
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||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || Miniprep || AraC || CBD test construct (pBad)
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || Minipreped || AraC || CBD test construct (pBad)


|- 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 || Miniprep || acGFP, dsRFP || CBD final construct
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI regulated promoter || Minipreped || acGFP, dsRFP || CBD final construct


|- 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 || Miniprep || --- || ''Not being used anymore''
||  || [http://parts.mit.edu/registry/index.php/Part:BBa_J07037 BBa_J07037] || cI and GFP || Minipreped || --- || ''Not being used anymore''


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Revision as of 03:51, 12 August 2007

Constructs Overview

Cloning Table

I Part Description Status Ligation Target Use in Constructs
BBa_E0040 GFP Minipreped --- Not being used anymore
BBa_I13522 ptet-GFP Minipreped --- Basic construct for chassis characterization
BBa_J5528 pBad-GFP Not Cloned --- Test construct for CBD (pBad)
I BBa_J24677 ptet-luxR-plux-GFP Minipreped --- Not being used anymore
BBa_R0040 ptet Minipreped acGFP, dsRFP, luxI CBD final (pBad), ID AHL sender, Hrp test construct
I BBa_I0500 pBad Minipreped acGFP, dsRFP CBD final construct, Hrp test construct
BBa_F2620 ptet-luxR-plux Minipreped lacZ, Hrp amplifier ID final construct
BBa_F1610 luxI Minipreped ptet ID AHL sender final construct
BBa_R0010 lacI-pLac Minipreped --- Not being used anymore
BBa_E1010 RFP Minipreped --- Not being used anymore
BBa_E7104 T7 GFP Minipreped --- CBD test construct (T7)
BBa_S03550 RBS-AraC-noStop Minipreped ptet, pBad CBD final construct (pBad)
BBa_B0015 Double terminator Minipreped AraC CBD test construct (pBad)
BBa_R0051 λcI regulated promoter Minipreped acGFP, dsRFP CBD final construct
BBa_J07037 cI and GFP Minipreped --- Not being used anymore
BBa_E0240 GFP Not cloned cI CBD test construct (cI)
BBa_I13507 RFP Not cloned CBD promoters CBD final reporter
BBa_T9002 ptet-luxR-plux-GFP Not cloned --- Basic construct for chassis characterization
BBa_I719??? T7 promoter Pending acGFP, dsRFP CBD final construct
BBa_I719??? acGFP Ordered ptet, pBad, plux Reporter system for test constructs, Hrp
BBa_I719??? dsRFP Ordered ptet, pBad, plux Reporter system for ID, Hrp
BBa_I719??? HrpR-HrpS-pHrpL Ordered Promoter, acGFP Hrp core system
BBa_I719??? HrpV Ordered Promoter, dsRFP Hrp inhibitor system

Ligation Table

I Part Description Status Parents Application
BBa_I719??? ptet-dsRFP Pending BBa_I13522+dsRFP CBD final construct (ptet)
BBa_I719??? pBad-dsRFP Pending BBa_I0500+dsRFP CBD final construct (pBad)
BBa_I719??? ptet-luxI Pending R0040+F1610 ID AHL sender final construct

Legend: I (Information)

Parts from the Registry

A W G S Name Description Plasmid Resist Copy Size(B) Size(T) Location
A W G BBa_E0040 GFP pSB1A2 AmpR High 720bp 2799bp Well 5H, Plate 1
A W G BBa_I13522 GFP under tetracyclin promoter pSB1A2 AmpR High 937bp 3016bp Well 15H, Plate 2
A BBa_J5528 GFP under arabinose promoter pSB2K3 KanR Low 2093bp 6518bp Well 6H, Plate 3
A W G BBa_J24677 GFP under lux promoter, with luxR pSB1A2 AmpR High 1787bp 3866bp Well 18K, Plate 3
A W X BBa_R0040 tetracyclin promoter pSB1A2 AmpR High 54bp 2133bp Well 7O, Plate 1
A W G BBa_I0500 arabinose promoter pSB2K3 KanR Low 1210bp 5635bp Well 9I, Plate 2
A W G BBa_F2620 lux promoter, with luxR pSB1A2 AmpR High 1061bp 3140bp Well 6B, Plate 1
A W G BBa_F1610 luxI coding region pSB1AK3 A+KR High 798bp 3987bp Well 1B, Plate 2
A W G BBa_R0010 lac promoter pSB1A2 AmpR High 200bp 2279bp Well 7K, Plate 1
A G BBa_E1010 RFP pSB2K3 KanR Low 681bp 5106bp Well 15M, Plate 2
A W G BBa_E7104 T7 GFP pSB1A2 AmpR High 826bp 2905bp Well 13F, Plate 2
A G BBa_S03550 AraC with RBS but no stop pSB1AC3 A+CR High 958bp 4013bp Well 20E, Plate 3
A W G BBa_B0015 Double terminator pSB1AK3 A+KR High 129bp 3316bp Well 1B, Plate 2
A W X BBa_R0051 λcI promoter pSB1A2 AmpR High 49bp 2128bp Well 9C, Plate 1
A G BBa_J07037 GFP under cI promoter pSB2K3 KanR Low 912bp 5337bp Well 12D, Plate 2
A BBa_E0240 GFP pSB1A2 AmpR High 876bp 2955bp Well 16A, Plate 1
A BBa_I13507 RFP pSB1A2 AmpR High 861bp 2940bp Well 16N, Plate 1
A W BBa_T9002 GFP under lux promoter, with luxR pSB1A3 AmpR High 1945bp 4102bp Well 19B, Plate 2

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 Name Description Plasmid Parents Resist Copy Size Stock Project


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


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
PHrpL promoting GFP


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