pGEX-5G/LIC

名称	编号	规格	价格	手册
pGEX-5G/LIC

质粒编号:
质粒名称: pGEX-5G/LIC
其他名称:
质粒大小（bp）: 4968
抗性: Ampicillin
质粒类型: Unspecified
是否病毒: Unspecified
启动子:
稳转或瞬转: Unspecified
组成型或诱导性: Unspecified
表达水平:
蛋白标签:
测序引物:
GenBank号: M97937
其他属性: The order of the major features in this plasmid is:pMB1 ori – lacIq – lacZ’ – tac – GST/SalI/SacII/thrombin cleavage site/BamHI – ampR. Vector is covered by a U.S. patent application and is distributed only for non-commercial use. (personal communication) Ligation-independent expression vector for constructing fusion proteins with a carrier polypeptide (glutathione S-transferase) which allows single-step affinity purification. The carrier protein can be removed by thrombin proteolysis. Preparation of the vector for cloning includes linearization with SacII, gel purification of the linearized vector, and treatment with T4 DNA polymerase in the presence of dATP. Target sequences for cloning are prepared by PCR and do not require restriction enzyme digestion. Design of PCR primers permits cloning in any reading frame. The forward primer should contain 15 nt complementary to nt 5′ to the SacII site of the vector [(5′-3′)(GGCCTGGTTCCGCGG)] followed by 12-15 nt corresponding to the target sequence (nt encoding the N-terminal 4-5 aa of the protein). The reverse primer should contain 14 nt complementary to nt 3′ to the SacII site of the vector [(5′-3′) (CTGCGCCTCGCTCC)] followed by 12 nt complementary to the 3′ end of the target sequence. Annealing the vector and amplification product forms a gapped duplex molecule that can be used directly to transform bacteria without ligation. Both primers should contain a dAMP residue near the sequence complementary to the vector to terminate the exonucleolytic activity of subsequent T4 DNA polymerase treatment. Restriction digests of the clone give the following sizes (kb): SacII–5.1; PstI–5.1; SalI–5.1; PstI/BamHI–4.0, 0.98. (ATCC staff) Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, E.coli, E.coli DH5alpha
质粒图谱:

质粒序列:

1	acgttatcga	ctgcacggtg	caccaatgct	tctggcgtca	ggcagccatc	50
51	ggaagctgtg	gtatggctgt	gcaggtcgta	aatcactgca	taattcgtgt	100
101	cgctcaaggc	gcactcccgt	tctggataat	gttttttgcg	ccgacatcat	150
151	aacggttctg	gcaaatattc	tgaaatgagc	tgttgacaat	taatcatcgg	200
201	ctcgtataat	gtgtggaatt	gtgagcggat	aacaatttca	cacaggaaac	250
251	agtattcatg	tcccctatac	taggttattg	gaaaattaag	ggccttgtgc	300
301	aacccactcg	acttcttttg	gaatatcttg	aagaaaaata	tgaagagcat	350
351	ttgtatgagc	gcgatgaagg	tgataaatgg	cgaaacaaaa	agtttgaatt	400
401	gggtttggag	tttcccaatc	ttccttatta	tattgatggt	gatgttaaat	450
451	taacacagtc	tatggccatc	atacgttata	tagctgacaa	gcacaacatg	500
501	ttgggtggtt	gtccaaaaga	gcgtgcagag	atttcaatgc	ttgaaggagc	550
551	ggttttggat	attagatacg	gtgtttcgag	aattgcatat	agtaaagact	600
601	ttgaaactct	caaagttgat	tttcttagca	agctacctga	aatgctgaaa	650
651	atgttcgaag	atcgtttatg	tcataaaaca	tatttaaatg	gtgatcatgt	700
701	aacccatcct	gacttcatgt	tgtatgacgc	tcttgatgtt	gttttataca	750
751	tggacccaat	gtgcctggat	gcgttcccaa	aattagtttg	ttttaaaaaa	800
801	cgtattgaag	ctatcccaca	aattgataag	tacttgaaat	ccagcaagta	850
851	tatagcatgg	cctttgcagg	gctggcaagc	cacgtttggt	ggtgtcgacc	900
901	atcctccagg	aggaggagga	ggcctggttc	cgcggagcga	ggcgcagttg	950
951	gatccccggg	aattcatcgt	gactgactga	cgatctgcct	cgcgcgtttc	1000
1001	ggtgatgacg	gtgaaaacct	ctgacacatg	cagctcccgg	agacggtcac	1050
1051	agcttgtctg	taagcggatg	ccgggagcag	acaagcccgt	cagggcgcgt	1100
1101	cagcgggtgt	tggcgggtgt	cggggcgcag	ccatgaccca	gtcacgtagc	1150
1151	gatagcggag	tgtataattc	ttgaagacga	aagggcctcg	tgatacgcct	1200
1201	atttttatag	gttaatgtca	tgataataat	ggtttcttag	acgtcaggtg	1250
1251	gcacttttcg	gggaaatgtg	cgcggaaccc	ctatttgttt	atttttctaa	1300
1301	atacattcaa	atatgtatcc	gctcatgaga	caataaccct	gataaatgct	1350
1351	tcaataatat	tgaaaaagga	agagtatgag	tattcaacat	ttccgtgtcg	1400
1401	cccttattcc	cttttttgcg	gcattttgcc	ttcctgtttt	tgctcaccca	1450
1451	gaaacgctgg	tgaaagtaaa	agatgctgaa	gatcagttgg	gtgcacgagt	1500
1501	gggttacatc	gaactggatc	tcaacagcgg	taagatcctt	gagagttttc	1550
1551	gccccgaaga	acgttttcca	atgatgagca	cttttaaagt	tctgctatgt	1600
1601	ggcgcggtat	tatcccgtgt	tgacgccggg	caagagcaac	tcggtcgccg	1650
1651	catacactat	tctcagaatg	acttggttga	gtactcacca	gtcacagaaa	1700
1701	agcatcttac	ggatggcatg	acagtaagag	aattatgcag	tgctgccata	1750
1751	accatgagtg	ataacactgc	ggccaactta	cttctgacaa	cgatcggagg	1800
1801	accgaaggag	ctaaccgctt	ttttgcacaa	catgggggat	catgtaactc	1850
1851	gccttgatcg	ttgggaaccg	gagctgaatg	aagccatacc	aaacgacgag	1900
1901	cgtgacacca	cgatgcctgc	agcaatggca	acaacgttgc	gcaaactatt	1950
1951	aactggcgaa	ctacttactc	tagcttcccg	gcaacaatta	atagactgga	2000
2001	tggaggcgga	taaagttgca	ggaccacttc	tgcgctcggc	ccttccggct	2050
2051	ggctggttta	ttgctgataa	atctggagcc	ggtgagcgtg	ggtctcgcgg	2100
2101	tatcattgca	gcactggggc	cagatggtaa	gccctcccgt	atcgtagtta	2150
2151	tctacacgac	ggggagtcag	gcaactatgg	atgaacgaaa	tagacagatc	2200
2201	gctgagatag	gtgcctcact	gattaagcat	tggtaactgt	cagaccaagt	2250
2251	ttactcatat	atactttaga	ttgatttaaa	acttcatttt	taatttaaaa	2300
2301	ggatctaggt	gaagatcctt	tttgataatc	tcatgaccaa	aatcccttaa	2350
2351	cgtgagtttt	cgttccactg	agcgtcagac	cccgtagaaa	agatcaaagg	2400
2401	atcttcttga	gatccttttt	ttctgcgcgt	aatctgctgc	ttgcaaacaa	2450
2451	aaaaaccacc	gctaccagcg	gtggtttgtt	tgccggatca	agagctacca	2500
2501	actctttttc	cgaaggtaac	tggcttcagc	agagcgcaga	taccaaatac	2550
2551	tgtccttcta	gtgtagccgt	agttaggcca	ccacttcaag	aactctgtag	2600
2601	caccgcctac	atacctcgct	ctgctaatcc	tgttaccagt	ggctgctgcc	2650
2651	agtggcgata	agtcgtgtct	taccgggttg	gactcaagac	gatagttacc	2700
2701	ggataaggcg	cagcggtcgg	gctgaacggg	gggttcgtgc	acacagccca	2750
2751	gcttggagcg	aacgacctac	accgaactga	gatacctaca	gcgtgagcta	2800
2801	tgagaaagcg	ccacgcttcc	cgaagggaga	aaggcggaca	ggtatccggt	2850
2851	aagcggcagg	gtcggaacag	gagagcgcac	gagggagctt	ccagggggaa	2900
2901	acgcctggta	tctttatagt	cctgtcgggt	ttcgccacct	ctgacttgag	2950
2951	cgtcgatttt	tgtgatgctc	gtcagggggg	cggagcctat	ggaaaaacgc	3000
3001	cagcaacgcg	gcctttttac	ggttcctggc	cttttgctgg	ccttttgctc	3050
3051	acatgttctt	tcctgcgtta	tcccctgatt	ctgtggataa	ccgtattacc	3100
3101	gcctttgagt	gagctgatac	cgctcgccgc	agccgaacga	ccgagcgcag	3150
3151	cgagtcagtg	agcgaggaag	cggaagagcg	cctgatgcgg	tattttctcc	3200
3201	ttacgcatct	gtgcggtatt	tcacaccgca	taaattccga	caccatcgaa	3250
3251	tggtgcaaaa	cctttcgcgg	tatggcatga	tagcgcccgg	aagagagtca	3300
3301	attcagggtg	gtgaatgtga	aaccagtaac	gttatacgat	gtcgcagagt	3350
3351	atgccggtgt	ctcttatcag	accgtttccc	gcgtggtgaa	ccaggccagc	3400
3401	cacgtttctg	cgaaaacgcg	ggaaaaagtg	gaagcggcga	tggcggagct	3450
3451	gaattacatt	cccaaccgcg	tggcacaaca	actggcgggc	aaacagtcgt	3500
3501	tgctgattgg	cgttgccacc	tccagtctgg	ccctgcacgc	gccgtcgcaa	3550
3551	attgtcgcgg	cgattaaatc	tcgcgccgat	caactgggtg	ccagcgtggt	3600
3601	ggtgtcgatg	gtagaacgaa	gcggcgtcga	agcctgtaaa	gcggcggtgc	3650
3651	acaatcttct	cgcgcaacgc	gtcagtgggc	tgatcattaa	ctatccgctg	3700
3701	gatgaccagg	atgccattgc	tgtggaagct	gcctgcacta	atgttccggc	3750
3751	gttatttctt	gatgtctctg	accagacacc	catcaacagt	attattttct	3800
3801	cccatgaaga	cggtacgcga	ctgggcgtgg	agcatctggt	cgcattgggt	3850
3851	caccagcaaa	tcgcgctgtt	agcgggccca	ttaagttctg	tctcggcgcg	3900
3901	tctgcgtctg	gctggctggc	ataaatatct	cactcgcaat	caaattcagc	3950
3951	cgatagcgga	acgggaaggc	gactggagtg	ccatgtccgg	ttttcaacaa	4000
4001	accatgcaaa	tgctgaatga	gggcatcgtt	cccactgcga	tgctggttgc	4050
4051	caacgatcag	atggcgctgg	gcgcaatgcg	cgccattacc	gagtccgggc	4100
4101	tgcgcgttgg	tgcggatatc	tcggtagtgg	gatacgacga	taccgaagac	4150
4151	agctcatgtt	atatcccgcc	gtcaaccacc	atcaaacagg	attttcgcct	4200
4201	gctggggcaa	accagcgtgg	accgcttgct	gcaactctct	cagggccagg	4250
4251	cggtgaaggg	caatcagctg	ttgcccgtct	cactggtgaa	aagaaaaacc	4300
4301	accctggcgc	ccaatacgca	aaccgcctct	ccccgcgcgt	tggccgattc	4350
4351	attaatgcag	ctggcacgac	aggtttcccg	actggaaagc	gggcagtgag	4400
4401	cgcaacgcaa	ttaatgtgag	ttagctcact	cattaggcac	cccaggcttt	4450
4451	acactttatg	cttccggctc	gtatgttgtg	tggaattgtg	agcggataac	4500
4501	aatttcacac	aggaaacagc	tatgaccatg	attacggatt	cactggccgt	4550
4551	cgttttacaa	cgtcgtgact	gggaaaaccc	tggcgttacc	caacttaatc	4600
4601	gccttgcagc	acatccccct	ttcgccagct	ggcgtaatag	cgaagaggcc	4650
4651	cgcaccgatc	gcccttccca	acagttgcgc	agcctgaatg	gcgaatggcg	4700
4701	ctttgcctgg	tttccggcac	cagaagcggt	gccggaaagc	tggctggagt	4750
4751	gcgatcttcc	tgaggccgat	actgtcgtcg	tcccctcaaa	ctggcagatg	4800
4801	cacggttacg	atgcgcccat	ctacaccaac	gtaacctatc	ccattacggt	4850
4851	caatccgccg	tttgttccca	cggagaatcc	gacgggttgt	tactcgctca	4900
4901	catttaatgt	tgatgaaagc	tggctacagg	aaggccagac	gcgaattatt	4950
4951	tttgatggcg	ttggaatt	4968