IGEM:IMPERIAL/2007/DNA Constructs

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(Difference between revisions)

Revision as of 12:10, 31 August 2007

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	BBa_I13522	ptet-GFP	XL1, Midi, Mini, BL21	---	Basic construct for chassis characterization
	11	BBa_E7104	pT7 GFP	XL1, Midi, Mini, BL21	---	CBD test construct (pT7)
I	18	BBa_T9002	ptet-luxR-plux-GFP	XL1, Midi, BL21	---	Basic construct for chassis characterization

	05	BBa_R0040	ptet	XL1, Midi, Mini	DsRed	CBD final (ptet), ID AHL sender
		BBa_I719???	pT7	Pending	DsRed	CBD final construct (pT7)
	07	BBa_F2620	ptet-luxR-plux	XL1, Midi, Mini	DsRed	ID final construct
	14	BBa_R0051	pcI	Problematic	DsRed	CBD final construct (pcI)

	21	BBa_J23039	ptet-luxI	XL1, Midi	---	ID sender construct
	24	BBa_R0062	plux	XL1, Midi	---	ID construct 2
	25	BBa_J37032	plux-GFP	XL1, Midi	---	ID construct 2
	26	BBa_S03119	ptet-luxR	XL1, Midi	---	---

	27	pSB1A3	Empty pSB1A3	Waiting Cloning	---	---
	28	BBa_P1010	pSB1A3 with ccd	Waiting Cloning	---	---
	29	BBa_P1010	pSB1A2 with ccd	Waiting Cloning	---	---

	13	BBa_B0015	Double terminator	XL1, Midi, Mini	---	General part construction
	17	BBa_I13507	RFP	XL1, Midi	---	Not being used anymore
	19	BBa_B0034	strong RBS	XL1, Waiting Midi	---	General part construction
	23	BBa_I13504	GFP	XL1, Midi	pcI	General part construction

		BBa_I719???	DsRed	Ordered	ptet, plux, pcI	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_I719022	pcI & GFP	Paused	BBa_R0051 + BBa_I13504	CBD test construct (pcI)
		BBa_I719???	pT7 & GFP	Proposed	pT7 + BBa_I13504	CBD test construct (pT7)

		BBa_I719???	DsRed & pSB1A3	Planning	DsRed +BBa_P1010	Biobrick Formation

		BBa_I719???	pT7 & DsRed	Planning	pT7 + DsRed	CBD final construct (pT7)
		BBa_I719???	ptet & DsRed	Planning	BBa_R0040 + DsRed	CBD final construct (ptet)
		BBa_I719???	pcI-DsRed	Paused	BBa_R0051 + DsRed	CBD final construct (pcI)
		BBa_I719???	ptet-luxR-plux-DsRed	Pending	BBa_F2620 + DsRed	ID detector final construct 1
		BBa_I719???	plux-DsRed	Pending	BBa_R0062 + DsRed	ID detector final construct 2

Legend: I (Information), [Status: XL1 - cloned in XL1-Blue; Midi - midiprepped; Mini - miniprepped; BL21 - cloned in BL21(DE3)]

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

110/100

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

W

X

14

BBa_R0051

λcI promoter

pSB1A2

AmpR

High

49bp

2128bp

Well 9C, Plate 1

--

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

100

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

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

100

A

G

26

BBa_S03119

constitutive LuxR

pSB1A2

AmpR

High

998bp

3077bp

Well 19J, Plate 1

60

A

W

27

pSB1A3

pSB1A3 (blank)

pSB1A3

Amp

High

0bp

2157bp

Well 23E, Plate 1

--

A

W

28

BBa_P1010

pSB1A3 (ccd)

pSB1A3

Amp

High

675bp

2832bp

Well 20I, Plate 3

--

A

W

29

BBa_P1010

pSB1A2 (ccd)

pSB1A2

Amp

High

675bp

2754bp

Well 20G, Plate 3

--

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 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
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
		20	---	AraC	pSB1AC3	1095bp	Planned	araC construct abandoned

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

Availability: Found in registry as BBa_I13522

pT7 GFP

PT7 promoting GFP

Availability: Found in resigry as BBa_E7104

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

Availability: Found in registry as BBa_T9002

pLux GFP with LuxR

Plux promoting GFP

Availability: Found in registry as BBa_J37032

<<Two more constructs with DsRed express yet to be included>>

@@ Line 52: / Line 52: @@
 |- style="background:#CCFFFF" align="center"
 ||  || 26 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03119 BBa_S03119] || ptet-luxR || <font color=#CC0000>XL1</font>, <font color=#0000FF>'''Midi'''</font> || --- || ---
+|- style="background:#CCFFFF" align="center"
+||  ||  ||  ||  ||  ||  ||
+<!-- BRICKS WITH PLASMID VECTORS -->
+|- style="background:#CCFFFF" align="center"
+||  || 27 || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || Empty pSB1A3 || Waiting Cloning || --- || ---
+|- style="background:#CCFFFF" align="center"
+||  || 28 || [http://parts.mit.edu/registry/index.php/Part:BBa_P1010 BBa_P1010] || pSB1A3 with ccd || Waiting Cloning || --- || ---
+|- style="background:#CCFFFF" align="center"
+||  || 29 || [http://parts.mit.edu/registry/index.php/Part:BBa_P1010 BBa_P1010] || pSB1A2 with ccd || Waiting Cloning || --- || ---
 |- style="background:#CCFFFF" align="center"
@@ Line 75: / Line 87: @@
 |- style="background:#CCFFFF" align="center"
-||  ||  || BBa_I719??? || HrpR-HrpS-pHrpL || Ordered || ??? || Hrp regulators
+||  ||  || BBa_I719??? || HrpR-HrpS || Ordered || ??? || Hrp regulators
 |- style="background:#CCFFFF" align="center"
@@ Line 99: / Line 111: @@
 |width="--"%|Application
 |- style="background:#CCFFCC" align="center"
-||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I719021 BBa_I719021] || ptet & luxI || ''Deleted'' || [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_I719022 BBa_I719022] || pcI & GFP || ''Paused'' || [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] +  [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || CBD test construct (pcI)
 |- style="background:#CCFFCC" align="center"
-||  ||  || [http://parts.mit.edu/registry/index.php/Part:BBa_I719022 BBa_I719022] || pcI & GFP || ''Paused''|| [http://parts.mit.edu/registry/index.php/Part:BBa_R0051 BBa_R0051] +  [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || CBD test construct (pcI)
+||  ||  || BBa_I719??? || pT7 & GFP || Proposed || pT7 + [http://parts.mit.edu/registry/index.php/Part:BBa_I13504 BBa_I13504] || CBD test construct (pT7)
+|- style="background:#CCFFCC" align="center"
+||  ||  ||  ||  ||  ||  ||
+|- style="background:#CCFFCC" align="center"
+||  ||  || BBa_I719??? || DsRed & pSB1A3 || Planning || DsRed +[http://parts.mit.edu/registry/index.php/Part:BBa_P1010 BBa_P1010] || Biobrick Formation
 |- style="background:#CCFFCC" align="center"
@@ Line 187: / Line 205: @@
 |- style="background:#FFCCFF" align="center"
 || A ||  || G ||  || 26 || [http://parts.mit.edu/registry/index.php/Part:BBa_S03119 BBa_S03119] || constitutive LuxR || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || AmpR || High || 998bp || 3077bp || Well 19J, Plate 1 || 60
+|- style="background:#FFCCFF" align="center"
+|| A || W ||  ||  || 27 || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || pSB1A3 (blank) || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || Amp || High || 0bp || 2157bp || Well 23E, Plate 1 || --
+|- style="background:#FFCCFF" align="center"
+|| A || W ||  ||  || 28 || [http://parts.mit.edu/registry/index.php/Part:BBa_P1010  BBa_P1010] || pSB1A3 (ccd) || [http://parts.mit.edu/registry/index.php/Part:pSB1A3 pSB1A3] || Amp || High || 675bp || 2832bp || Well 20I, Plate 3 || --
+|- style="background:#FFCCFF" align="center"
+|| A || W ||  ||  || 29 || [http://parts.mit.edu/registry/index.php/Part:BBa_P1010  BBa_P1010] || pSB1A2 (ccd) || [http://parts.mit.edu/registry/index.php/Part:pSB1A2 pSB1A2] || Amp || High || 675bp || 2754bp || Well 20G, Plate 3 || --
 |- style="background:#FFCCFF" align="center"

IGEM:IMPERIAL/2007/DNA Constructs

From OpenWetWare

Revision as of 12:10, 31 August 2007

Contents

Constructs Overview

Cloning Table

Ligation Table

Parts from the Registry

Our Parts

Part Graveyard

Construct Diagrams

CBD Constructs

ID Constructs

Views

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Contributing Authors