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

Revision as of 05:57, 14 August 2007

Constructs Overview

Cloning Table

I	ID	Part	Description	Status	Ligation Target	Use in Constructs
	01	BBa_E0040	GFP	Minipreped	---	Not being used anymore
	02	BBa_I13522	ptet-GFP	Minipreped	---	Basic construct for chassis characterization
	03	BBa_J5528	pBad-GFP	Cloning	---	Test construct for CBD (pBad)
	05	BBa_R0040	ptet	Minipreped	acGFP, DsRed, luxI	CBD final (pBad), ID AHL sender, Hrp test construct
I	06	BBa_I0500	pBad	Minipreped	acGFP, DsRed	CBD final construct, Hrp test construct
	07	BBa_F2620	ptet-luxR-plux	Minipreped	lacZ, Hrp amplifier	ID final construct
	08	BBa_F1610	luxI	Minipreped	ptet	ID AHL sender final construct
	09	BBa_R0010	lacI-pLac	Minipreped	---	Not being used anymore
	10	BBa_E1010	RFP	Minipreped	---	Not being used anymore
	11	BBa_E7104	T7 GFP	Minipreped	---	CBD test construct (T7)
	12	BBa_S03550	RBS-AraC-noStop	Minipreped	ptet, pBad	CBD final construct (pBad)
	13	BBa_B0015	Double terminator	Minipreped	AraC	CBD test construct (pBad)
	14	BBa_R0051	λcI regulated promoter	Minipreped	acGFP, DsRed	CBD final construct
	16	BBa_E0240	GFP	Cloning	cI	CBD test construct (cI)
	17	BBa_I13507	RFP	Cloning	CBD promoters	CBD final reporter
I	18	BBa_T9002	ptet-luxR-plux-GFP	Cloning	---	Basic construct for chassis characterization
	19	BBa_B0034	strong RBS	Cloning	---	Backup part
		BBa_I719???	T7 promoter	Pending	acGFP, DsRed	CBD final construct
		BBa_I719???	acGFP	Ordered	ptet, pBad, plux	Reporter system for test constructs, Hrp
		BBa_I719???	DsRed	Ordered	ptet, pBad, plux	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
		BBa_I719???	lacZ	Stopped	---	Not being used anymore

Ligation Table

I	ID	Part	Description	Status	Parents	Application
		BBa_I719020	araC generator	Planned	BBa_S03550 + BBa_B0015	CBD test construct 3 and final constructs 5, 8 and 10
		BBa_I719021	ptet & luxI	Concerns	R0040 + F1610	ID final sender (duplicated)
		BBa_I719022	PcI & GFP	Pending	BBa_R0051 + BBa_E0240	CBD test construct 4
		BBa_I719???	Ptet & araC	Pending	BBa_R0040 + araC with stop	CBD test construct 3 and final construct 8
		BBa_I719???	Ptet-araC & pBad-GFP	Pending	Ptet-araC + BBa_J5528	CBD test construct 3
		BBa_I719???	PcI & araC	Pending	BBa_R0051 + araC with stop	CBD test construct 5 and final construct 10
		BBa_I719???	PcI-araC & pBad-GFP	Pending	PcI-araC + BBa_J5528	CBD test construct 5
		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???	pBad & DsRed	Pending	BBa_I0500 + DsRed	CBD final construct 8 and 10
		BBa_I719???	Ptet-araC & pBad-DsRed	Pending	Ptet-araC + pBad-DsRed	CBD final construct 8
		BBa_I719???	pcI-DsRed	Pending	BBa_R0051 + DsRed	CBD final construct 9
		BBa_I719???	PcI-araC & pBad-DsRed	Pending	PcI-araC + pBad-DsRed	CBD final construct 10
		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???	RBS & DsRed	Pending	BBa_B0034 + DsRed	ID detector final construct 4 and 5
		BBa_I719???	RBS-DsRed & stop	Pending	RBS-DsRed + BBa_B0015	ID detector final construct 4 and 5
		BBa_I719???	ptet-luxR-plux-DsRed	Pending	BBa_F2620 + DsRed (with RBS and stop)	ID detector final construct 4
		BBa_I719???	Ptet-luxR-Plux-hrpR-hrpS-phrpL & DsRed	Pending	Ptet-luxR-Plux-hrpR-hrpS-phrpL + DsRed (with RBS and stop)	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

03

BBa_J5528

GFP under arabinose promoter

pSB2K3

KanR

Low

2093bp

6518bp

Well 6H, Plate 3

A

W

X

05

BBa_R0040

tetracyclin promoter

pSB1A2

AmpR

High

54bp

2133bp

Well 7O, Plate 1

A

W

G

06

BBa_I0500

arabinose promoter

pSB2K3

KanR

Low

1210bp

5635bp

Well 9I, Plate 2

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

09

BBa_R0010

lac promoter

pSB1A2

AmpR

High

200bp

2279bp

Well 7K, Plate 1

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

G

12

BBa_S03550

AraC with RBS but no stop

pSB1AC3

A+CR

High

958bp

4013bp

Well 20E, Plate 3

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

16

BBa_E0240

GFP

pSB1A2

AmpR

High

876bp

2955bp

Well 16A, 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

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_I791???

luxI coding region under tetracyclin promoter

pSB1A2

AmpR

High

860bp

2939bp

BBa_I719020

AraC protein generator

pSB1AC3?

AmpR

High

933bp

3988bp

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		04	BBa_J24677	plux-GFP	pSB1A2	1787bp	Miniprepped	Wrong GFP ligated
A		15	BBa_J07037	cI-GFP	pSB2K3	912bp	Miniprepped	No RBS present on this part

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 13: / Line 13: @@
 |- 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''
-<!-- TETRACYCLIN PROMOTER WITH GFP-->
 |- 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
@@ Line 19: / Line 19: @@
 |- style="background:#CCFFFF" align="center"
 ||  || 03 || [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || pBad-GFP || Cloning || --- || Test construct for CBD (pBad)
-|- style="background:#CCFFFF" align="center"
-|| [[IGEM:IMPERIAL/2007/DNA_Constructs/BBa_J24677|I]] || 04 || [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] || ptet-luxR-plux-GFP || Minipreped || --- || ''Not being used anymore''
 |- style="background:#CCFFFF" align="center"
@@ Line 60: / Line 57: @@
 |- style="background:#CCFFFF" align="center"
-||  || 18 || [http://parts.mit.edu/registry/index.php/Part:BBa_T9002 BBa_T9002] || ptet-luxR-plux-GFP || Cloning || --- || Basic construct for chassis characterization
+|| [[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
 |- style="background:#CCFFFF" align="center"
@@ Line 139: / Line 136: @@
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || hrpR-hrpS & phrpL|| Pending ||hrpR-hrpS + phrpL || ID AHL detector test construct 3 and final construct 5
+||  ||  || BBa_I719??? || hrpR-hrpS & phrpL|| Pending ||hrpR-hrpS + phrpL || ID detector test construct 3 and final construct 5
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || Ptet-luxR-Plux & hrpR-hrpS-phrpL || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + hrpR-hrpS-phrpL || ID AHL detector test construct 3 and final construct 5
+||  ||  || BBa_I719??? || Ptet-luxR-Plux & hrpR-hrpS-phrpL || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + hrpR-hrpS-phrpL || ID detector test construct 3 and final construct 5
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || Ptet-luxR-Plux-hrpR-hrpS-phrpL & GFP || Pending ||Ptet-luxR-Plux-hrpR-hrpS-phrpL + [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || ID AHL detector test construct 3
+||  ||  || BBa_I719??? || Ptet-luxR-Plux-hrpR-hrpS-phrpL & GFP || Pending ||Ptet-luxR-Plux-hrpR-hrpS-phrpL + [http://parts.mit.edu/registry/index.php/Part:BBa_E0240 BBa_E0240] || ID detector test construct 3
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || RBS & DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] + DsRed || ID AHL detector final construct 4 and 5
+||  ||  || BBa_I719??? || RBS & DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_B0034 BBa_B0034] + DsRed || ID detector final construct 4 and 5
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || RBS-DsRed & stop || Pending || RBS-DsRed + [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || ID AHL detector final construct 4 and 5
+||  ||  || BBa_I719??? || RBS-DsRed & stop || Pending || RBS-DsRed + [http://parts.mit.edu/registry/index.php/Part:BBa_B0015 BBa_B0015] || ID detector final construct 4 and 5
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || ptet-luxR-plux-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + DsRed (with RBS and stop)|| ID AHL detector final construct 4
+||  ||  || BBa_I719??? || ptet-luxR-plux-DsRed || Pending || [http://parts.mit.edu/registry/index.php/Part:BBa_F2620 BBa_F2620] + DsRed (with RBS and stop)|| ID detector final construct 4
 |- style="background:#CCFFCC" align="center"
-||  ||  || BBa_I719??? || Ptet-luxR-Plux-hrpR-hrpS-phrpL & DsRed|| Pending ||Ptet-luxR-Plux-hrpR-hrpS-phrpL + DsRed (with RBS and stop) || ID AHL detector final construct 5
+||  ||  || BBa_I719??? || Ptet-luxR-Plux-hrpR-hrpS-phrpL & DsRed|| Pending ||Ptet-luxR-Plux-hrpR-hrpS-phrpL + DsRed (with RBS and stop) || ID detector final construct 5
 |- style="background:#CCFFCC" align="center"
@@ Line 190: / Line 187: @@
 || A ||  ||  ||  || 03 || [http://parts.mit.edu/registry/index.php/Part:BBa_J5528 BBa_J5528] || GFP under arabinose promoter || [http://parts.mit.edu/registry/index.php/Part:pSB2K3 pSB2K3] || KanR || Low || 2093bp || 6518bp || Well 6H, Plate 3
-|- style="background:#FFCCFF" align="center"
-|| A || W || G ||  || 04 || [http://parts.mit.edu/registry/index.php/Part:BBa_J24677 BBa_J24677] || GFP under lux promoter, with luxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 1787bp || 3866bp || Well 18K, Plate 3
-<!-- Number 05 -->
 |- style="background:#FFCCFF" align="center"
 || A || W || <font color=#990033>X</font> ||  || 05 || [http://parts.mit.edu/registry/index.php/Part:BBa_R0040 BBa_R0040] || tetracyclin promoter || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High ||  54bp || 2133bp || Well 7O, Plate 1
@@ Line 285: / Line 279: @@
 |width="--"%| Notes
 |- style="background:#DDDDDD" align="center"
-|| A ||  || 15 || BBa_J07037 || cI-GFP || pSB2K3 || 912bp || Minipreped || No RBS present on this part
+|| A ||  || 04 || BBa_J24677 || plux-GFP || pSB1A2 || 1787bp || Miniprepped || Wrong GFP ligated
+|- 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"
-||  ||  ||  ||  ||  ||  ||  ||
+||  ||  ||  ||  ||  ||  ||  ||  ||
 |}

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

Revision as of 05:57, 14 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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