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

Revision as of 10:56, 15 August 2007

Constructs Overview

Cloning Table

I	ID	Part	Description	Status	Ligation Target	Use in Constructs
	02	BBa_I13522	ptet-GFP	Midipreped	---	Basic construct for chassis characterization
	11	BBa_E7104	T7 GFP	Midipreped	---	CBD test construct (T7)
I	18	BBa_T9002	ptet-luxR-plux-GFP	Midiprepped	---	Basic construct for chassis characterization
	22	BBa_S03335	cI-GFP	Not Cloned	---	CBD test construct (cI)

	05	BBa_R0040	ptet	Midipreped	acGFP, DsRed	CBD final (pBad), ID AHL sender, Hrp test construct
		BBa_I719???	T7 promoter	Pending	acGFP, DsRed	CBD final construct
	07	BBa_F2620	ptet-luxR-plux	Midipreped	DsRed	ID final construct
	14	BBa_R0051	λcI regulated promoter	Midipreped	acGFP, DsRed	CBD final construct

	08	BBa_F1610	luxI	Midipreped	ptet	ID AHL sender final construct
	21	BBa_J23039	ptet-LuxI	Not Cloned	---	ID sender construct

	01	BBa_E0040	GFP	Minipreped	---	Not being used anymore
	10	BBa_E1010	RFP	Minipreped	---	Not being used anymore
	13	BBa_B0015	Double terminator	Midipreped	---	General part construction
	17	BBa_I13507	RFP	Midiprepped	---	Not being used anymore
	19	BBa_B0034	strong RBS	Waiting Midi	---	General part construction

		BBa_I719???	acGFP	Ordered	ptet, plux, cI	Reporter system for test constructs, Hrp
		BBa_I719???	DsRed	Ordered	ptet, plux, cI	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

Ligation Table

I	ID	Part	Description	Status	Parents	Application
		BBa_I719021	ptet & luxI	Concerns	R0040 + F1610	ID final sender (duplicated - J23039)
		BBa_I719022	cI & GFP	Planned	BBa_R0051 + BBa_E0240	CBD test construct 4
		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???	pcI-DsRed	Pending	BBa_R0051 + DsRed	CBD final construct 9
		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-DsRed	Pending	BBa_F2620 + DsRed	ID detector final construct 4
		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)

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

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

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

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

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_I719022

cI with GFP

pSB1A2

AmpR

High

933bp

3988bp

BBa_I791021

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		03	BBa_J5528	pBad-GFP	pSB2K3	2093bp	Failed	ara construct abandoned
A		04	BBa_J24677	plux-GFP	pSB1A2	1787bp	Miniprepped	Wrong GFP ligated
A		06	BBa_I0500	pBad	pSB2K3	1210bp	Midiprepped	ara construct abandoned
A	W	08	BBa_F1610	luxI	pSB1AK3	798bp	Midiprepped	Substitute part J23039
A	W	09	BBa_R0010	lacI-plac	pSB1A2	200bp	Miniprepped	Lac construct abandoned
A		12	BBa_S03550	araC	pSB1AC3	958bp	Midiprepped	araC construct abandoned
A		15	BBa_J07037	cI-GFP	pSB2K3	912bp	Miniprepped	No RBS present on this part
A	W	16	BBa_E0240	GFP	pSB1A2	876bp	Midiprepped	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.

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.

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.

Construct CBD 5
- All parts are currently available in the Biobricks Registry.
- pcI promoter: BBa_R0051 (available), in pSB1A2 (AmpR)
- Arabinose C regulator protein with RBS: BBa_S03550 (available), in pSB1AC3 (A+CR)
- Stop codon: BBa_B0015, in pSB1AK3 (A+KR)
- pBAD with GFP: BBa_J5528, in pSB2K3 (KanR)

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 promoter avaiable in the registry: BBa_I0500
- dsRed has been ordered
- Arabinose C regulator protein with RBS: BBa_S03550 (available), in pSB1AC3 (A+CR)
- Stop codon: BBa_B0015, in pSB1AK3 (A+KR)
- tet promoter available in the registry: BBa_R0040

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.

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.

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

@@ Line 10: / Line 10: @@
 |width=22%|Ligation Target
 |width="--"%|Use in Constructs
-<!-- T7 PROMOTER -->
+|- style="background:#CCFFFF" align="center"
+||  || 02 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || Midipreped || --- || Basic construct for chassis characterization
 |- style="background:#CCFFFF" align="center"
-||  || 01 || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || Minipreped || --- || ''Not being used anymore''
+||  || 11 || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP || Midipreped || --- || CBD test construct (T7)
+|- style="background:#CCFFFF" align="center"
+|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || 18 || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || Midiprepped || --- || Basic construct for chassis characterization
 |- style="background:#CCFFFF" align="center"
-||  || 02 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13522 BBa_I13522] || ptet-GFP || Minipreped || --- || Basic construct for chassis characterization
+||  || 22 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03335 BBa_S03335] || cI-GFP || Not Cloned || --- || CBD test construct (cI)
 |- style="background:#CCFFFF" align="center"
-||  || 05 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || Minipreped || acGFP, DsRed, luxI || CBD final (pBad), ID AHL sender, Hrp test construct
+||  ||  ||  ||  ||  ||  ||
 |- style="background:#CCFFFF" align="center"
-||  || 07 || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || Minipreped || lacZ, Hrp amplifier || ID final construct
+||  || 05 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || ptet || Midipreped || acGFP, DsRed || CBD final (pBad), ID AHL sender, Hrp test construct
 |- style="background:#CCFFFF" align="center"
-||  || 08 || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI || Minipreped || ptet || ID AHL sender final construct
+||  ||  || BBa_I719??? || T7 promoter || Pending || acGFP, DsRed || CBD final construct
 |- style="background:#CCFFFF" align="center"
-||  || 10 || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || Minipreped || --- || ''Not being used anymore''
+||  || 07 || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] || ptet-luxR-plux || Midipreped || DsRed || ID final construct
 |- style="background:#CCFFFF" align="center"
-||  || 11 || [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 BBa_E7104] || T7 GFP || Minipreped || --- || CBD test construct (T7)
+||  || 14 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI regulated promoter || Midipreped || acGFP, DsRed || CBD final construct
 |- style="background:#CCFFFF" align="center"
-||  || 13 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || Minipreped || AraC || CBD test construct (pBad)
+||  ||  ||  ||  ||  ||  ||
 |- style="background:#CCFFFF" align="center"
-||  || 14 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI regulated promoter || Minipreped || acGFP, DsRed || CBD final construct
+||  || 08 || [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 BBa_F1610] || luxI || Midipreped || ptet || ID AHL sender final construct
 |- style="background:#CCFFFF" align="center"
-||  || 16 || [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || GFP || Cloning || cI || CBD test construct (cI)
+||  || 21 || [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 BBa_J23039] || ptet-LuxI || Not Cloned || --- || ID sender construct
 |- style="background:#CCFFFF" align="center"
-||  || 17 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || Cloning || CBD promoters || CBD final reporter
+||  ||  ||  ||  ||  ||  ||
 |- style="background:#CCFFFF" align="center"
-|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || 18 || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || Cloning || --- || Basic construct for chassis characterization
+||  || 01 || [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0040 BBa_E0040] || GFP || Minipreped || --- || ''Not being used anymore''
 |- style="background:#CCFFFF" align="center"
-||  || 19 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || strong RBS || Cloning || --- || Backup part
+||  || 10 || [http://parts.mit.edu/registry/index.php/Part:BBa_E1010 BBa_E1010] || RFP || Minipreped || --- || ''Not being used anymore''
 |- style="background:#CCFFFF" align="center"
-||  || 21 || [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 BBa_J23039] || ptet-LuxI || Not Cloned || --- || ID sender construct
+||  || 13 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || Double terminator || Midipreped || --- || ''General part construction''
+|- style="background:#CCFFFF" align="center"
+||  || 17 || [http://parts.mit.edu/registry/index.php/Part:BBa_I13507 BBa_I13507] || RFP || Midiprepped || --- || ''Not being used anymore''
+|- style="background:#CCFFFF" align="center"
+||  || 19 || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] || strong RBS || Waiting Midi || --- || ''General part construction''
 |- style="background:#CCFFFF" align="center"
-||  ||  || BBa_I719??? || T7 promoter || Pending || acGFP, DsRed || CBD final construct
+||  ||  ||  ||  ||  ||  ||
 |- style="background:#CCFFFF" align="center"
-||  ||  || BBa_I719??? || acGFP || Ordered || ptet, pBad, plux || Reporter system for test constructs, Hrp
+||  ||  || BBa_I719??? || acGFP || Ordered || ptet, plux, cI || Reporter system for test constructs, Hrp
 |- style="background:#CCFFFF" align="center"
-||  ||  || BBa_I719??? || DsRed || Ordered || ptet, pBad, plux || Reporter system for ID, Hrp
+||  ||  || BBa_I719??? || DsRed || Ordered || ptet, plux, cI || Reporter system for ID, Hrp
 |- style="background:#CCFFFF" align="center"
@@ Line 66: / Line 78: @@
 |- style="background:#CCFFFF" align="center"
-||  ||  || BBa_I719??? || HrpV|| Ordered || Promoter, DsRed || Hrp inhibitor system
+||  ||  || BBa_I719??? || HrpV || Ordered || Promoter, DsRed || Hrp inhibitor system
 |- style="background:#CCFFFF" align="center"
@@ Line 83: / Line 95: @@
 |width="--"%|Application
 |- style="background:#CCFFCC" align="center"
-||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I719021 BBa_I719021] || cI & luxI || ''Concerns'' || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 R0040] + [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 F1610] || ID final sender (''duplicated'' - [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 J23039])
+||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I719021 BBa_I719021] || ptet & luxI || ''Concerns'' || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 R0040] + [http://parts.mit.edu/registry/index.php/Part:BBa_F1610 F1610] || ID final sender (''duplicated'' - [http://parts.mit.edu/registry/index.php/Part:BBa_J23039 J23039])
 |- style="background:#CCFFCC" align="center"
@@ Line 160: / Line 172: @@
 |- style="background:#FFCCFF" align="center"
 || A || W || <font color=#990033>X</font> ||  || 14 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] || &lambda;cI promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 49bp || 2128bp || Well 9C, Plate 1
-|- style="background:#FFCCFF" align="center"
-|| A ||  ||  ||  || 16 || [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || GFP || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 876bp || 2955bp || Well 16A, Plate 1
 |- style="background:#FFCCFF" align="center"
@@ Line 233: / Line 242: @@
 |- style="background:#DDDDDD" align="center"
 || A ||  || 06 || BBa_I0500 || pBad || pSB2K3 || 1210bp || Midiprepped|| ara construct abandoned
+|- style="background:#DDDDDD" align="center"
+|| A || W || 08 || BBa_F1610 || luxI || pSB1AK3 || 798bp || Midiprepped || Substitute part J23039
 |- style="background:#DDDDDD" align="center"
@@ Line 242: / Line 254: @@
 |- style="background:#DDDDDD" align="center"
 || A ||  || 15 || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || Miniprepped || No RBS present on this part
+|- style="background:#DDDDDD" align="center"
+|| A || W || 16 || BBa_E0240 || GFP || pSB1A2 || 876bp || Midiprepped || Redundant
 |- style="background:#DDDDDD" align="center"

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

Revision as of 10:56, 15 August 2007

Contents

Constructs Overview

Cloning Table

Ligation Table

Parts from the Registry

Our Parts

Part Graveyard

Constructs for Cell By Date

Constructs for Infector Detector

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