IGEM:IMPERIAL/2007/DNA Constructs

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Revision as of 03:20, 14 August 2007 by Ben Yi Tew (talk | contribs)
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Constructs Overview

Cloning Table

I ID Part Description Status Ligation Target Use in Constructs
01 BBa_E0040 GFP Minipreped --- Not being used anymore
02 BBa_I13522 ptet-GFP Minipreped --- Basic construct for chassis characterization
03 BBa_J5528 pBad-GFP Cloning --- Test construct for CBD (pBad)
I 04 BBa_J24677 ptet-luxR-plux-GFP Minipreped --- Not being used anymore
05 BBa_R0040 ptet Minipreped acGFP, DsRed, luxI CBD final (pBad), ID AHL sender, Hrp test construct
I 06 BBa_I0500 pBad Minipreped acGFP, DsRed CBD final construct, Hrp test construct
07 BBa_F2620 ptet-luxR-plux Minipreped lacZ, Hrp amplifier ID final construct
08 BBa_F1610 luxI Minipreped ptet ID AHL sender final construct
09 BBa_R0010 lacI-pLac Minipreped --- Not being used anymore
10 BBa_E1010 RFP Minipreped --- Not being used anymore
11 BBa_E7104 T7 GFP Minipreped --- CBD test construct (T7)
12 BBa_S03550 RBS-AraC-noStop Minipreped ptet, pBad CBD final construct (pBad)
13 BBa_B0015 Double terminator Minipreped AraC CBD test construct (pBad)
14 BBa_R0051 λcI regulated promoter Minipreped acGFP, DsRed CBD final construct
16 BBa_E0240 GFP Cloning cI CBD test construct (cI)
17 BBa_I13507 RFP Cloning CBD promoters CBD final reporter
18 BBa_T9002 ptet-luxR-plux-GFP Cloning --- Basic construct for chassis characterization
19 BBa_B0034 strong RBS Cloning --- Backup part
BBa_I719??? T7 promoter Pending acGFP, DsRed CBD final construct
BBa_I719??? acGFP Ordered ptet, pBad, plux Reporter system for test constructs, Hrp
BBa_I719??? DsRed 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, DsRed Hrp inhibitor system
BBa_I719??? lacZ Stopped --- Not being used anymore

Ligation Table

I ID Part Description Status Parents Application


BBa_I719??? RBS-araC & stop codon Pending BBa_S03550 + BBa_B0015 CBD test construct 3 and final constructs 5, 8 and 10
BBa_I719??? Ptet & araC Pending BBa_R0040 + araC with stop CBD test construct 3 and final construct 8
BBa_I719??? Ptet-araC & pBad-gfp Pending Ptet-araC + BBa_J5528 CBD test construct 3
BBa_I719??? PcI & gfp Pending BBa_R0051 + BBa_E0240 CBD test construct 4
BBa_I719??? PcI & araC Pending BBa_R0051 + araC with stop CBD test construct 5 and final construct 10
BBa_I719??? PcI-araC & pBad-gfp Pending PcI-araC + BBa_J5528 CBD test construct 5
BBa_I719??? pT7 & DsRed Pending pT7 + DsRed CBD final construct 6
BBa_I719??? ptet & DsRed Pending BBa_R0040+ DsRed CBD final construct 7
BBa_I719??? pBad & DsRed Pending BBa_I0500 + DsRed CBD final construct 8 and 10
BBa_I719??? Ptet-araC & pBad-DsRed Pending Ptet-araC + pBad-DsRed CBD final construct 8
BBa_I719??? pcI-DsRed Pending BBa_R0051 + DsRed CBD final construct 9
BBa_I719??? PcI-araC & pBad-DsRed Pending PcI-araC + pBad-DsRed CBD final construct 10
BBa_I719??? ptet & luxI Pending R0040 + F1610 ID AHL sender final construct 1
BBa_I719??? hrpR & hrpS Pending hrpR + hrpS ID AHL detector test construct 3 and final construct 5 (already got this??)
BBa_I719??? hrpR-hrpS & phrpL Pending hrpR-hrpS + phrpL ID AHL detector test construct 3 and final construct 5
BBa_I719??? Ptet-luxR-Plux & hrpR-hrpS-phrpL Pending BBa_F2620 + hrpR-hrpS-phrpL ID AHL 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 AHL detector test construct 3
BBa_I719??? RBS & DsRed Pending BBa_B0034 + DsRed ID AHL detector final construct 4 and 5
BBa_I719??? RBS-DsRed & stop Pending RBS-DsRed + BBa_B0015 ID AHL detector final construct 4 and 5
BBa_I719??? ptet-luxR-plux-DsRed Pending BBa_F2620 + DsRed (with RBS and stop) ID AHL detector final construct 4
BBa_I719??? Ptet-luxR-Plux-hrpR-hrpS-phrpL & DsRed Pending Ptet-luxR-Plux-hrpR-hrpS-phrpL + DsRed (with RBS and stop) ID AHL 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 03 BBa_J5528 GFP under arabinose promoter pSB2K3 KanR Low 2093bp 6518bp Well 6H, Plate 3
A W G 04 BBa_J24677 GFP under lux promoter, with luxR pSB1A2 AmpR High 1787bp 3866bp Well 18K, Plate 3
A W X 05 BBa_R0040 tetracyclin promoter pSB1A2 AmpR High 54bp 2133bp Well 7O, Plate 1
A W G 06 BBa_I0500 arabinose promoter pSB2K3 KanR Low 1210bp 5635bp Well 9I, Plate 2
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 09 BBa_R0010 lac promoter pSB1A2 AmpR High 200bp 2279bp Well 7K, Plate 1
A G 10 BBa_E1010 RFP pSB2K3 KanR Low 681bp 5106bp Well 15M, Plate 2
A W G 11 BBa_E7104 T7 GFP pSB1A2 AmpR High 826bp 2905bp Well 13F, Plate 2
A G 12 BBa_S03550 AraC with RBS but no stop pSB1AC3 A+CR High 958bp 4013bp Well 20E, Plate 3
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 16 BBa_E0240 GFP pSB1A2 AmpR High 876bp 2955bp Well 16A, 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

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_I791??? 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 15 BBa_J07037 cI-GFP pSB2K3 912bp Miniprepped No RBS present on this part

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
    • Parts avaiable ....
pT7 promoting RFP


  • Construct CBD 7
    • Parts avaiable ....
Ptet promoting RFP


  • Construct CBD 8
    • Parts avaiable ....
PBAD promoting RFP and Ptet promoting araC


  • Construct CBD 9
    • Parts avaiable ....
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
    • Parts available ...
Ptet promoting LuxI


  • Construct ID 5
    • Parts available ...
Ptet promoting LuxR, Plux promoting HrpR and HrpS and PHrpL promoting lacZ


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