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

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||  ||  || BBa_I719??? || pcI-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] + DsRed || CBD final construct (pcI)
||  ||  || BBa_I719??? || pcI-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] + DsRed || CBD final construct (pcI)
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|| A ||  ||  ||  || 22 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03335 BBa_S03335] || GFP under cI promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 932bp || 3011bp || ''Well 85, Box 9?'' || --
 
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Revision as of 03:12, 16 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 pT7 GFP XL1, Midi, Mini, BL21 --- CBD test construct (pT7)
I 18 BBa_T9002 ptet-luxR-plux-GFP XL1, Midi --- Basic construct for chassis characterization
22 BBa_S03335 pcI-GFP Not Cloned --- CBD test construct (pcI)
05 BBa_R0040 ptet XL1, Midi, Mini DsRed CBD final (ptet), ID AHL sender
BBa_I719??? pT7 Pending DsRed CBD final construct (pT7)
07 BBa_F2620 ptet-luxR-plux XL1, Midi, Mini DsRed ID final construct
14 BBa_R0051 pcI XL1, Midi, Mini DsRed CBD final construct (pcI)
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, pcI Reporter system for test constructs, Hrp
BBa_I719??? DsRed Ordered ptet, plux, pcI 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 pcI & GFP Concerns BBa_R0051 + BBa_E0240 CBD test construct (pcI)
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), [Status: XL1 - cloned in XL1-Blue; Midi - midiprepped; Mini - miniprepped; BL21 - cloned in BL21(DE3)]

Parts from the Registry

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(T) Location Conc.
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 --
A 22 BBa_S03335 GFP under cI promoter pSB1A2 AmpR High 932bp 3011bp Well 85, Box 9? --

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 ptet pSB1A2 AmpR High 860bp 2939bp
BBa_I791??? DsRed under ptet pSB1A2 AmpR High

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

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 Substitute part I13507
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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