Team:Harvard/Plasmids

From 2008.igem.org

(Difference between revisions)

Revision as of 02:52, 27 October 2008

Parts

[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet]

Internal name	Description	Registry Name	Part Size	Vector (size)	Plasmid size (vector + part)	Origin	Marker	Transformed?	Miniprep	Glycerol Stock	Sequenced?
P1	Low promoter+RBS +GFP+term	J23113/I20259	919	pSB3K3 (2750)	3669	p15A	Kan	in S1 and TOP10	Yes for both strains	Yes for both strains.	S1P1A_A, P1E2 (tube labeled E1A)
P2	Med promoter+RBS+ GFP+term	J23150/I20269	919	pSB3K3 (2750)	3669	p15A	Kan	in TOP10, not S1	Yes		P2E2 (tube labeled 2B)
P3	High promoter+RBS+ GFP+term	J23151/I20270	919	pSB3K3 (2750)	3669	p15A	Kan	in TOP10 and S1	Yes for both strains.	Yes for both strains.	P3E2 (tube labeled E3B), S1P3A_A
P4	Low-Medium copy vector	pACYC duet	4008			p15a	Cm	in S1 and TOP10	Yes, for both strains.	Yes for both strains.
P5	Low-Medium copy vector (w/ death gene)	pSB3K3	2750			p15a	Kan	in E3	Yes	Yes (B- done 6/30. A-done 7/1)
P11	Light responsive system, dual regulation	BBa_M30109	4333	pSB1AC3 (3055)	7388	pMB1	Amp/Cm	Failed
P13	Low copy plasmid	pCDFDuet-1	3781			CloDF13	Sm	in S1	Yes, 435 ng/uL	Yes
P15	pTet+RBS+GFP+term	BBa_I13522	937	BBa_J23006 (3279)		ColE1	Amp	in E2	Yes	No	P15A, P15B
P16	Tet repressible promoter	BBa_R0040	54			pMB1	Amp	in E2	Yes	No
P17	Inverter (TetR w/o promoter and Tet promoter)	BBa_Q04400	902	pSB2K3 (4425)	5327	OriS, P1 lytic, F1	Kan	in E1	BAD	BAD	cells from MIT correct
P18	cI regulated promoter	R0051	49			pMB1	Amp	in E2	Yes	Yes, 7/4	P18A, P18B appear so (20 bp align at end, but ref is 49 bp, so this isn\'t bad)
P20	GFP (LVA tagged) with Lac promoter	BBa_J04431	1122			pMB1	Amp, Kan	in E2	Yes	No	?! P20A: ~100 bp of beginning of F read aligned to ref; R read did not
P21	pACYC (P4) w/ biobricks	A1 (pACYC)	???			???	???	in S1 and TOP10	Yes	Yes, for both strains.
P22	ompR controlled promoter for ompC (long)	BBa_R0082	108			pMB1	Amp				WRONGWRONGWRONG: DID NOT ALIGN TO REF
P23	ompR controlled promoter for ompC (short)	BBa_R0083	78			pMB1	Amp	in E2	Yes	No	WRONGWRONGWRONG: DID NOT ALIGN TO REF
P24	ompR controlled promoter for ompF	BBa_R0084	108			pMB1	Amp	in E2	Yes	No	WRONGWRONGWRONG: DID NOT ALIGN TO REF
P25	Promoter activated by OmpR-P with the reporter GFP	BBa_J58102	991			pMB1	Amp				WRONGWRONGWRONG: DID NOT ALIGN TO REF
P26	Mutated promoter activated by OmpR-P with the reporter GFP	BBa_J58103	961			pMB1	Amp	in E2	Yes	No	WRONGWRONGWRONG: DID NOT ALIGN TO REF
P27	repressor plasmid w/ constitutive expression of LacI and TetR	pZS4int2				pSC101	Sm	Yes	Yes (6/26)	Yes (6/26)
P28	GFP w/ PLtctO-1	pZE21				ColE1	Kan	Yes	Yes (6/26)	Yes (6/26)
P29	Venus (mutant YFP) PL (Plambda)	pZS*2R				pSC101*	Kan	Yes	Yes (6/26)	Yes (6/26)
P30	YFP w/ PLlacO-1	pZA32				p15A	Cm	Yes	Yes (6/26)	Yes (6/26)
P31	Venus w/ PLlacO-1	pZS*1R				pSC101*	Amp	Yes	Yes (6/26)	Yes (6/26)
P32	Venus w/ PLlacO-1	pZA32				p15A	Cm	Yes	Yes (6/26)	Yes (6/26)
P33	Venus w/ PLtctO-1	pZA31				p15A	Cm	Yes	Yes (6/26)	Yes (6/26)
P34	GFP w/ PLlacO-1	pZE12				ColE1	Amp	Yes	Yes (6/26)	Yes (6/26)
P35	accessory	pLDR8						Yes	Yes (6/26)	Yes (6/26)
P36	same as P20	BBa_I763004					Kan/Amp
P37	Constructive expression LacI w/ pTetR promoter	BBa_I722007					Amp	Failed (liquid cultures didn\'t grow)
P38	High constitutive promoter	BBa_J23114	35				Amp	2007 in E1 (DH5a), 2008 Failed, MIT	Yes BUT funny digestion pattern	2007 (B- done 6/30. A- done 7/1)
P39	Low constitutive promoter	BBa_J23113	35				Amp	2008 (A grew), 2007 (A grew), MIT	Yes BUT funny digestion pattern	2008 (A- done 6/30) 2007 (A-done 6/30)
P40	Ribosome Binding Site	BBa_B0032	13				Amp	2008 (A grew), 2007 (B grew)	Yes.	2007 (B -done 6/30) 2008 (A-done 6/30)
P41	Transcriptional Terminator	BBa_B0010					Amp	2007 (A and B grew, 7/2), 2008 (B grew, 7/2)		2007 (A and B done 7/4), 2008 (B done 7/4)
P42	LacI coding region	\"BBa_C0012
\"	1128				Amp	Failed (liquid cultures didn\'t grow)
P43	TetR coding region	BBa_C0040	660				Amp	2007 (A grew)	Yes	2007 (A-done 6/30)	P43A E1 07
P44	cI lambda	\"BBa_C0051
\"	750				Amp	2007 (A grew)	Yes	2007 (A-done 6/30)	P44A E1 07
P45	GFP only w/ RBS & terminator	BBa_E0240	876	pSB1A2 (2079)	2955	pMB1	Amp	2007 (A and B grew)	Yes	2007 (A and B -done 6/30)	P45A, P45B >E1 07 for both
P46	Base Vector	BBa_I51020					Amp	A and B grew.	Yes	A and B -done 6/30
P47	Kan Resistance Cassette	BBa_P1003	967				Kan	2007 (A and B grew, but spilled)	Yes	2007 (A and B -done 7/1)	P47A, P47B >E1 07 for both
P48	Cm Resistance Cassette	BBa_P1004	769	pSB1A1 (2708)	3477		Cm	2007 (A and B grew, but spilled)	Yes	2007 (A and B -done 7/1). Duplicate done 7/4.	P48B E1 07
P51	LacI QPI w/ RBS & promoter	BBa_Q04121	1370	pSB2K3 (4425)	5795	pMB1	Kan				cells from MIT correct
P52	cI QPI w/ RBS & promoter	BBa_Q04510	987	pSB2K3 (4425)	5412	pMB1	Kan
P53	PoPS --> TetR	BBa_P0440	840				Amp	2007 (B grew), 2008 (A grew)	Yes. (7/3).	2007 (B done 7/3), 2008 (A done 7/3)
P54	PoPS --> LacI	BBa_P0412	1308				Amp	2007 (A grew)	Yes. (7/3).	2007 (A done 7/3)
P55	RBS.Lambda.cI. LVA.Terminator	BBa_P0455	896				A/C	2007 (A and B grew). 2008 (A grew)	Yes for A and B (7/3).	2007 (A and B done 7/3). 2008 (A grew)
P56		RBS+TetR coding region (P40 and P43)					Amp	Yes in E1.	Yes for A and B (7/3).	A and B done 7/3
P57	RBS+lambda cI coding region (P40 and P44)	Composite					Amp	Yes in E1.	Yes for A and B (7/3).	A and B done 7/3
P58	GFP under pTet (with RBS and term) on plasmid with p15A origin (P1 vector + P15 part)	Composite	3016				Kan	Yes in E1.	Yes for A and B (7/3).	A and B done 7/3
P59	GFP under pLac (with RBS and term) on plasmid with p15A origin (P1 vector + P20 part)	Composite	3201				Kan	Yes in E1.	Yes for A and B (7/3).	A and B done 7/3
P60		Composite
P61	Terminator	BBa_B0011						2008 (A grew.)	Yes for 2008 A.	2008 (A done 7/4)
P62	Terminator	BBa_B0012						2007 (A and B grew). 2008 ((Plate yielded 1 colony only- failed to grow)	Yes for A and B 2007.	2007 (A and B done 7/4)
P63	Double terminator (P62 and P61)	BBa_B0014	95	pSB1AK3 (3189)	3284		Amp/Kan	2007 (A and B grew). 2008 (Plate yielded 1 colony only- failed to grow)	Yes for A and B 2007.	2007 (A and B done 7/4)
P64	Double terminator (P60 and P62)	BBa_B0015						2007 (A grew)		2007 (A done 7/4)
P65	Double terminator (P62 and P60)	BBa_B0025						Failed (liquid cultures didn\'t grow)
P66	hi promoter+RBS+TetR coding region (P38+P56)	Composite
P67	Low promoter+RBS+TetR	Composite
P68	hi promoter +RBS+lambda cI coding region (P38+P57)	Composite
P69	Low promoter+RBS+lambda cI repressor	Composite
P70	P66 and term	Composite
P71	P67 and term	Composite
P72	P68 and term	Composite
P73	P69 and term	Composite
P74	lambda cI promoter+RBS+GFP+term (P45 + P18)	Composite						Yes.	yes
P75	P74 + P1	Composite						yes in both S1 and E1	yes 7/22
P76	P17+P45	Tet QPI+GFP	1778	pSB1A2 (2079)	3857	pMB1	Amp	Yes	Yes	Yes
P77	RBS + LacI + terminators	BBa_P0312	1307					Failed (liquid cultures didn\'t grow)
P78	RBS + LacI + terminators	BBa_P0112	1310					Failed (liquid cultures didn\'t grow)
P79	P90+ P48	Composite
P80	P90 + P49	Composite						no- wrong
P83	P45+mtrA (w/ Pst1)	Composite
P84	heat sensitive Lac- YFP	J06912	2314	pSB1A2 (2079)		pMB1	Amp	Yes, in E1.	Yes, 7/9
P85	heat sensitive Lac-YFP	J06911	2314	pSB1A2 (2079)		pMB1	Amp	Yes, in E1.	yes
P86	pPL-PCB from UT Austin	n/a				P15a	Amp	Yes in S1
P87	pPL-cph1-envZ-pprotet	n/a				ColE1	Cm	No
P88	light-sensing EnvZ	BBa_I15010	2238	pSB2K3 (4425)		F\', P1 lytic	Kan	failed
P89	RBS + LacI + terminators + pLac	BBa_Q01121	1372	pSB1A2 (2079)	3451	pMB1	Kan	failed
P90	CDF in P1 vector			pSB3K3 (2750)		p15A	Kan
P91	P89+P45: lac QPI+complete GFP		4327				Amp
P92	P84 + P1	Composite						Q04510
P93	P85+P1	Composite
P94	P40 + mtrB	Composite	>4000	pSB1A2 (2079)			Amp
P95	RBS + cph8	BBa_S03419							Yes
P96	RBS + cph8 + terminators	BBa_S03417							Yes
P97	Strong RBS	BBa_B0034	12	pSB1A2 (2079)	2.1 kb	pMB1	Amp	Yes	Yes
P99	cell death	BBa_P1010									Part from MIT aligned to ref
P101	pSB4C5 (with death gene)	pSB4C5/I52002	1090	pSB4C5 (3221)	4311	pSC101	Cm	Yes	Yes	Yes	Part from MIT aligned to ref
P102	P108+P45 (lac induced GFP) in p15a vector		2281	pSB3K3 (2750)	5031	P15A	Kan	Yes	Yes	Yes	Yes, along with 108+45#41
P103	P108+P45 (lac induced GFP) in pSC101 vector		2281	pSB4C5 (3221)	5502	pSC101	Cm	Yes	Yes	Yes	P103 # 41, 45
P104	P75 + P63							Yes 1 colony	yes 7/28
P105	P87 w/o PstI site (mutated Cph)		2235	2022	4257	ColE1	Cm	yes			E2P105A, E5P105D
p106	pER21 - plasmid for genetic recombination	n/a : from Dad					Gentamycin	Yes	Yes
p107	pKOV - plasmid for genetic recombination	n/a: from George Church					Cm	Yes	Yes
P108	P3 + (P39+51) beta (lac QPI)	composite	1405	pSB3K3 (2750)	4155	p15A	Kan	yes	yes	yes	Verified
P109	RBS+full lacZ	I732017	3093	pSB1A2 (2079)	5172	pMB1	Amp	yes	yes	yes	correct from MIT
P110	RBS+lacZalpha+term	E0435	534	pSB4K5 (3419)	3953	pSC101	Kan	yes	yes	yes	\"correct from MIT
\"
P111	RBS+lacZalpha+term	E0435	534	pSB4C5 (3221)	3755	pSC101	Cm	yes	yes	yes	correct from MIT
P112	RBS+lacZalpha+term	E0435	534	pSB1AK3 ( 3189)	3723	pMB1	A/K	yes	yes	yes	correct from MIT
P113	RBS+lacZalpha+term	E0435	534	pSB4A3 (3339)	3873	repA pSC101	Amp	yes	yes	yes	correct from MIT
P114	pJBC2 (inducible IPTG system)								Yes
P115	low promoter+RBS+E. coli ompR+term (from alt method)		960	pSB3K3 (2750)	3710	P15A	Kan	Yes	Yes		ompR+P1 #64, 73 correct
P116	P38+P17 in pSB3K3		937	pSB3K3 (2750)	3687	P15A	Kan	Yes	Yes		P38+17 in P1/3 #2 , 3, 5
P117	P39+P17 in pSB3K3		937	pSB3K3 (2750)	3687	P15A	Kan	Yes	Yes		P39+17 in P1/3 #1
P118	low promoter+RBS+HO-pcyA+term in pSB3K3		1555	pSB3K3 (2750)	4305	P15A	Kan	Yes	Yes		HO+P1 #3
P119	P63+P18	Composite
P120	P63 + P18 + cI (RBS primer set)	Composite									verified (8/13)
P121	P104 + cI (RBS primer set)										verified (8/13)
P122	P120 + P38
P123	P120 + P39
P124	P121 + P38
P125	P121 + P39
P126	P115 (low ompR) in pSB4C5		960	pSB4C5 (3221)	4181	pSC101	Cm	Yes	Yes	Yes	Yes, along with 101+115 numbers 4, 14, 25
P127	P118 (low HO-pcyA) in pSB4C5		1555	pSB4C5 (3221)	4776	pSC101	Cm	Yes	Yes	Yes	Yes, along with 101+118 numbers 61, 77
P128	P97+mtrB		2106	pSB1A2 (2079)	4185	pMB1	Amp	Yes	Yes	Yes	128# 1, 5, 9, appear to have a silent SNP
P129	mtrB+P63		2189	pSB1AK3 (3189)	5378		Amp/Kan	Yes	Yes	Yes	129# 21, 28 appear to have a silent SNP
P130	P97+mtrB+P63	Composite	2201	pSB1AK3 (3189)	5390		Amp/Kan	Yes
P131	P130 in pSB4C5		2201	pSB4C5 (3221)	5422	pSC101	Cm				A and B have F>S
P132	P108 in pSB4C5		1405	pSB4C5 (3221)	4626	pSC101	Cm				A, D correct
P133	P116 in pSB4C5		937	pSB4C5 (3221)	4158	pSC101	Cm				A, B correct
P134	P117 in pSB4C5		937	pSB4C5 (3221)	4158	pSC101	Cm				A, B correct
P135	P39+P130		2236	pSB1A2 (2079)	4315	pMB1	Amp				#31, 39, 60, 70, 80 F>S, but need more checking for SNPs in mtrB
P136	P38+P130		2236	pSB1A2 (2079)	4315	pMB1	Amp				#20 F>S, but needs checking for SNPs in mtrB
P137	P135 in pSB4C5		2236	pSB4C5 (3221)	5457	pSC101	Cm				#32- HAS MUTATION IN MIDDLE, F>S
P138	P136 in pSB4C5										#1, 2> check for mutations in mtrB, are they not on the same vector?
P139	P130 in P133 (high constitutive promoter+tet QPI+RBS+mtrB+terminator)							no
P140	P108 in P131 (low constitutive promoter+lac QPI+RBS+mtrB+terminator		3637 (definite)	pSB4C5 (3221)	6858	pSC101	Cm				YES, but there is a F>S change in mtrB
P141	P38+51 (high promoter+lac QPI)		1405	pSB1A2 (2079)	3484	pMB1	Amp				#3, 4 correct
P142	P141 in P131 (high promoter+lac QPI+RBS+mtrB+terminator)		3637	pSB4C5 (3221)	6858	pSC101	Cm				YES but needs scrutiny (can\'t check that hi promoter is there, and SNPs in mtrB)
P143	P130 in P134 (low promoter+tet QPI+RBS+mtrB+term)							no
P144	high mtrB in 15a	38+130+1	2236	pSB3K3 (2750)	4986	p15A	Kan	yes	Yes	Yes
P145	low mtrB in 15a	39+130+1	2236	pSB3K3 (2750)	4986	p15A	Kan	yes	Yes	Yes
P146	high lacQPI+ mtrB in 15a	38+51+130+1	3637	pSB3K3 (2750)	6387	p15A	Kan	yes	Yes	Yes
P147	low lacQPI+ mtrB in 15a	39+51+130+1	3637	pSB3K3 (2750)	6387	p15A	Kan	yes	Yes	Yes

@@ Line 1: / Line 1: @@
-== Vectors ==
-*[[Media:PACYC-map.pdf‎|pACYC-Duet vector map]]
-*[http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3]
 ==Parts==
 *[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet]