pGEX-4T-2
PVT0030 2ug
pGEX-4T-2 Information
Promoter: Tac/lac
Replicon: ColE1 ori
Plasmid classification: Escherichia coli vector; PGEX series expression plasmid
Plasmid size: 4970bp
Plasmid label: N-GST, N-thrombin
Prokaryotic resistance: ampicillin Amp
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: pGEX5: GGGCTGGCAAGCCACGTTTGGTG
3'sequencing primers: pGEX3: CCGGGAGCTGCATGTGTCAGAGG
Note: the recombinant protein can be purified by GST affinity column
Use:pGEX Plasmid
pGEX-4T-2 Sequence
LOCUS Exported File 4970 bp ds-DNA circular SYN 28-1-2015
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4970)
TITLE Direct Submission
JOURNAL Exported 2015-1-28 from SnapGene 2.0.1
FEATURES Location/Qualifiers
source 1..4970
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 183..211
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac?UV5 promoters"
protein_bind 219..235
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
CDS 258..911
/codon_start=1
/gene="
"
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 918..935
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
promoter 1273..1377
/gene="bla"
/note="AmpR promoter"
CDS 1378..2238
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2409..2997
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3241..3318
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 3319..4401
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
protein_bind 4414..4435
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4450..4480
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4488..4504
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 4512..4528
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4544..4560)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
901 atcctccaaa atcggatctg gttccgcgtg gatccccagg aattcccggg tcgactcgag
961 cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac
1021 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
1081 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
1141 cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc
1201 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
1261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
1321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
1381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
1441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
1501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
1561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
1621 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
1681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
1741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
1801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
1861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
1921 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
1981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
2041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
2101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
2161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
2221 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
2281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
2341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
2401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
2461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
2521 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
2581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
2641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
2701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
2761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
2821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
2881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
2941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
3001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
3061 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
3121 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
3181 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc
3241 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
3301 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
3361 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
3421 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
3481 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
3541 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
3601 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
3661 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
3721 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
3781 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
3841 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
3901 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
3961 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
4021 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
4081 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
4141 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
4201 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
4261 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
4321 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
4381 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
4441 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
4501 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
4561 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
4621 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
4681 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
4741 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
4801 tgcacggtta cgatgcgccc atctacacca acgtaaccta tcccattacg gtcaatccgc
4861 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
4921 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt
//
Caution:
Product is for research use only!
Search name
pGEX-4T-2,Plasmid pGEX-4T-2,pGEX-4T-2 vector
English