Imperial/Wet Lab/DNA Constructs

From 2007.igem.org

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<!-- PROMOTERS WITH GFP -->
<!-- PROMOTERS WITH GFP -->
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||  || 02 || [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] || ptet-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || Basic construct for chassis characterization
+
||  || 02 || [http://partsregistry.org/Part:BBa_I13522 BBa_I13522] || ptet-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || [[Imperial/Cell_by_Date/Design#DNA_Constructs|Cell By Date concept 1]]
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
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||  || 11 || [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] || pT7 GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF><small>''Maxi''</small></font>,  <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || CBD test construct (pT7)
+
||  || 11 || [http://partsregistry.org/Part:BBa_E7104 BBa_E7104] || pT7 GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF><small>''Maxi''</small></font>,  <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font> || --- || [[Imperial/Cell_by_Date/Design#DNA_Constructs|Cell By Date concept 2]]
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||  || 30 || --- || pT7 (strong GFP) ||  <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font>|| --- || T7 test-construct
+
||  || 30 || --- || pT7 (strong GFP) ||  <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#00CC00>''Mini''</font>, <font color=#FF6633>BL21</font>|| --- || Construct to test T7 promoter
|- style="background:#CCFFFF" align="center"
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|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || 18 || [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF><small>''Midi''</small></font>, <font color=#FF6633>BL21</font> || --- || Basic construct for chassis characterization
+
|| || 18 || [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF><small>''Midi''</small></font>, <font color=#FF6633>BL21</font> || --- || [[Imperial/Infector_Detector/Design#DNA_Constructs|Infector Detector construct 1]]
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<!-- PROMOTERS ONLY -->
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||  || 05 || [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] || ptet || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || CBD final (ptet), ID AHL sender
+
||  || 05 || [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] || ptet || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || [[Imperial/Cell_by_Date/Design#DNA_Constructs|Cell By Date concept 1]] with visual reporter
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
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||  ||  || BBa_I719??? || pT7 || ''Ready'' || DsRedEx || CBD final construct (pT7)
+
||  ||  || [http://partsregistry.org/Part:BBa_I719005 BBa_I719005] || pT7 || <font color=#0000FF>''Midi''</font> || DsRedEx || [[Imperial/Cell_by_Date/Design#DNA_Constructs|Cell By Date concept 2]]
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|- style="background:#CCFFFF" align="center"
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||  || 07 || [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || ID final construct
+
||  || 07 || [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font>, <font color=#00CC00>''Mini''</font> || DsRedEx || [[Imperial/Infector_Detector/Design#DNA_Constructs|Infector Detector construct 1]] with visual reporter
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<!-- ID SPECIFIC BRICKS -->
<!-- ID SPECIFIC BRICKS -->
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||  || 21 || [http://partsregistry.org/Part:BBa_J23039 BBa_J23039] || ptet-luxI || <font color=#CC0000>XL1</font>, <font color=#0000FF><small>''Midi''</small></font> || --- || ID sender construct
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||  || 21 || [http://partsregistry.org/Part:BBa_J23039 BBa_J23039] || ptet-luxI || <font color=#CC0000>XL1</font>, <font color=#0000FF><small>''Midi''</small></font> || --- || AHL sender construct
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
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||  || 24 || [http://partsregistry.org/Part:BBa_R0062 BBa_R0062] || plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font> || DsRedEx || ID construct 2
+
||  || 24 || [http://partsregistry.org/Part:BBa_R0062 BBa_R0062] || plux || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font> || DsRedEx || [[Imperial/Infector_Detector/Design#DNA_Constructs|Infector Detector construct 2]] with visual reporter
|- style="background:#CCFFFF" align="center"
|- style="background:#CCFFFF" align="center"
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||  || 25 || [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] || plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF>''Midi''</font> || --- || ID construct 2
+
||  || 25 || [http://partsregistry.org/Part:BBa_J37032 BBa_J37032] || plux-GFP || <font color=#CC0000>XL1</font>, <font color=#CC00FF>''Maxi''</font>, <font color=#0000FF>''Midi''</font> || --- || [[Imperial/Infector_Detector/Design#DNA_Constructs|Infector Detector construct 2]]
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|- style="background:#CCFFFF" align="center"
 
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||  || 26 || [http://partsregistry.org/Part:BBa_S03119 BBa_S03119] || ptet-luxR || <font color=#CC0000>XL1</font>, <font color=#0000FF>''Midi''</font> || --- || ---
 
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||  ||  ||  ||  ||  ||  ||
 
<!-- BRICKS WITH PLASMID VECTORS -->
<!-- BRICKS WITH PLASMID VECTORS -->
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<!-- there are none of there : (-->
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||  || 27 || [http://partsregistry.org/Part:pSB1A3 pSB1A3] || Empty pSB1A3 || '''Problematic''' || --- || ---
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|| A ||  || G ||  || 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
|| A ||  || G ||  || 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
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|| A ||  || G ||  || 26 || [http://partsregistry.org/Part:BBa_S03119 BBa_S03119] || constitutive LuxR || [http://partsregistry.org/Part:pSB1A2 pSB1A2] || AmpR || High || 998bp || 3077bp || Well 19J, Plate 1 || 60
 
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|- style="background:#FFCCFF" align="center"
 
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|| A || W || <font color=#990033>X</font> ||  || 27 || [http://partsregistry.org/Part:pSB1A3 pSB1A3] || pSB1A3 (blank) || [http://partsregistry.org/Part:pSB1A3 pSB1A3] || Amp || High || 0bp || 2157bp || Well 23E, Plate 1 || --
 
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|- style="background:#DDDDDD" align="center"
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||  || W || 20 || --- || AraC || pSB1AC3 || 1095bp || Planned || araC construct abandoned
||  || W || 20 || --- || AraC || pSB1AC3 || 1095bp || Planned || araC construct abandoned
 +
 +
|- style="background:#DDDDDD" align="center"
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|| A ||  || 26 || BBa_S03119 || ptet-LuxR || pSB1A2 || XL1, Midi }} 998bp || not used in design
 +
 +
|- style="background:#DDDDDD" align="center"
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|| A ||  || 27 || pSB1A3 || pSB1A3 || pSB1A3 || failed-XL1 || 2157bp || biobrick not present
|- style="background:#DDDDDD" align="center"
|- style="background:#DDDDDD" align="center"

Revision as of 06:51, 26 October 2007


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:

I ID Part Description Status Ligation Target Use in Constructs
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
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 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 + [http://partsregistry.org/Part:BBa_I13504 BBa_I13504] CBD test construct (pT7)
BBa_I719??? DsRedEx & pSB1A3 Paused DsRedEx + [http://partsregistry.org/Part:BBa_P1010 BBa_P1010] Biobrick Submission
BBa_I719??? pT7 & DsRedEx Planned pT7 + DsRedEx CBD final construct (pT7)
BBa_I719??? ptet & DsRedEx Planned [http://partsregistry.org/Part:BBa_R0040 BBa_R0040] + DsRedEx CBD final construct (ptet)
BBa_I719??? ptet-luxR-plux-DsRedEx Planned [http://partsregistry.org/Part:BBa_T2620 BBa_F2620] + DsRedEx ID detector final construct 1
BBa_I719??? plux-DsRedEx Planned [http://partsregistry.org/Part:BBa_R0062 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.
(ng/ul)
A W G 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 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 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 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 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 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:

A W G S ID Name Description Plasmid Resist Copy Size(B) Size(P) Stock
[http://partsregistry.org/Part:BBa_I719022 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 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
Retrieved from "http://2007.igem.org/wiki/index.php/Imperial/Wet_Lab/DNA_Constructs"