Imperial/Wet Lab/DNA Constructs

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Wet Lab: DNA Constructs

Constructs Overview

Here are several tables listing the bricks which we are cloning, constructing and using, with their relevant details.

Cloning Table

Status of biobricks cloned and their applications:
Legend: I (Information), [Status: XL1 - cloned in XL1-Blue; Maxi - maxiprepped; Midi - midiprepped; Mini - miniprepped; BL21 - cloned in BL21(DE3); status - stocks running low]

I ID Part Description Status Ligation Target Use in Constructs
I 02 [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] ptet-GFP XL1, Maxi, Mini, BL21 --- Cell By Date concept 1
11 [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] pT7 GFP XL1, Maxi, Midi, Mini, BL21 --- Cell By Date concept 2
30 --- pT7 (strong GFP) XL1, Maxi, Mini, BL21 --- Construct to test T7 promoter
I 18 [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] ptet-luxR-plux-GFP XL1, Maxi, Midi, BL21 --- Infector Detector construct 1
05 [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] ptet XL1, Midi, Mini DsRedEx Cell By Date concept 1 with visual reporter
[http://partsregistry.org/Part:BBa_I719005 BBa_I719005] pT7 Midi DsRedEx Cell By Date concept 2
07 [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] ptet-luxR-plux XL1, Midi, Mini DsRedEx Infector Detector construct 1 with visual reporter
21 [http://partsregistry.org/Part:BBa_J23039 BBa_J23039] ptet-luxI XL1, Midi --- AHL sender construct
24 [http://partsregistry.org/Part:BBa_R0062 BBa_R0062] plux XL1, Midi DsRedEx Infector Detector construct 2 with visual reporter
25 [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] plux-GFP XL1, Maxi, Midi --- Infector Detector construct 2
13 [http://partsregistry.org/Part:BBa_B0015 BBa_B0015] Double terminator XL1, Maxi, Midi, Mini --- General part construction
17 [http://partsregistry.org/Part:BBa_I13507 BBa_I13507] RFP XL1, Midi --- Not being used anymore
19 [http://partsregistry.org/Part:BBa_B0034 BBa_B0034] strong RBS XL1 --- General part construction
23 [http://partsregistry.org/Part:BBa_I13504 BBa_I13504] GFP XL1, Maxi, Midi pT7 Used to construct [http://partsregistry.org/Part:BBa_I719015 BBa_I719015]
BBa_I719??? DsRed Express Delayed ptet, plux, pT7 The Visual reporter
BBa_I719??? HrpR-HrpS Ready  ??? Hrp regulators
BBa_I719??? pHrpL Ready  ??? Hrp promoter
BBa_I719??? HrpV Ready  ??? Hrp inhibitor system


Ligation Table

Status of parts ligated and their applications:
Legend: I (Information), [Status: XL1 - cloned in XL1-Blue; Maxi - maxiprepped; Midi - midiprepped; Mini - miniprepped; BL21 - cloned in BL21(DE3); status - stocks running low]

I ID Part Description Status Parents Application
I [http://partsregistry.org/Part:BBa_I719015 BBa_I719015] pT7 & GFP XL1, Maxi, Mini, BL21 pT7 + [http://partsregistry.org/Part:BBa_I13504 BBa_I13504] Cell By Date concept 2
BBa_I719??? DsRedEx & pSB1A3 Paused DsRedEx + [http://partsregistry.org/Part:BBa_P1010 BBa_P1010] Biobrick Submission
BBa_I719??? pT7 & DsRedEx Planned pT7 + DsRedEx Cell By Date concept 2 with visual reporter
BBa_I719??? ptet & DsRedEx Planned [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] + DsRedEx Cell By Date concept 1 with visual reporter
BBa_I719??? ptet-luxR-plux-DsRedEx Planned [http://partsregistry.org/Part:BBa_T2620 BBa_F2620] + DsRedEx Infector Detector construct 1 with visual reporter
BBa_I719??? plux-DsRedEx Planned [http://partsregistry.org/Part:BBa_R0062 BBa_R0062] + DsRedEx Infector Detector construct 2 with visual reporter

Parts from the Registry

Physical information of biobricks:
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)

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(T) Location Conc.
(ng/ul)
A W G S 02 [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] GFP under tetracyclin promoter [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 937bp 3016bp Well 15H, Plate 2 500
A W G 05 [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] tetracyclin promoter [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 54bp 2133bp Well 7O, Plate 1 60
A W G S 07 [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] lux promoter, with luxR [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 1061bp 3140bp Well 6B, Plate 1 80
A X G S 11 [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] T7 GFP [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 826bp 2905bp Well 13F, Plate 2 100
A W G 13 [http://partsregistry.org/Part:BBa_B0015 BBa_B0015] Double terminator [http://partsregistry.org/Part:pSB1AK3 pSB1AK3] A+KR High 129bp 3316bp Well 1B, Plate 2 100/70
A G 17 [http://partsregistry.org/Part:BBa_I13507 BBa_I13507] RFP [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 861bp 2940bp Well 16N, Plate 1 80
A W G S 18 [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] GFP under lux promoter with LuxR [http://partsregistry.org/Part:pSB1A3 pSB1A3] AmpR High 1945bp 4102bp Well 19B, Plate 2 460
A W 19 [http://partsregistry.org/Part:BBa_B0034 BBa_B0034] RBS [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 12bp 2091bp Well 3O, Plate 1 --
A G 21 [http://partsregistry.org/Part:BBa_J23039 BBa_J23039] Constitutive luxI [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 860bp 2939bp Well 11K, Plate 3 140
A W G S 23 [http://partsregistry.org/Part:BBa_I13504 BBa_I13504] GFP [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 875bp 2954bp Well 16E, Plate 1 120/330
A G 24 [http://partsregistry.org/Part:BBa_R0062 BBa_R0062] lux promoter [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 55bp 2134bp Well 9G, Plate 1 90
A G S 25 [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] GFP under lux promoter [http://partsregistry.org/Part:pSB1A2 pSB1A2] AmpR High 938bp 3017bp Well 8D, Plate 3 125/330
W 30 --- T7 with GFP(strong) --- Kan?  ??? ---  ??? --- 150


Our Parts

Physical information of our ligated parts:
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

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(P) Stock
A W G S [http://partsregistry.org/Part:BBa_I791005 BBa_I791005] T7 promoter pSB1A2 AmpR High 23bp 2102bp
A W G S [http://partsregistry.org/Part:BBa_I791015 BBa_I791015] GFP under T7 promoter pSB1A2 AmpR High 906bp 2985bp

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 26 BBa_S03119 ptet-LuxR pSB1A2 XL1, Midi 998bp not used in design
A 27 pSB1A3 pSB1A3 pSB1A3 failed-XL1 2157bp biobrick not present
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
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