IGEM:IMPERIAL/2007/DNA Constructs

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<html><h3>THE IMPERIAL iGEM 2007 Team </h3></html>
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Fabrication & Testing Protocols

DNA Constructs
Overview of DNA Constructs

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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.
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 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>>