pGEX-KG Search name
pGEX-KG,Plasmid pGEX-KG,pGEX-KG vector
pGEX-KG Plasmid Information
Promoter: Tac/lac
Replicon: ColE1 ori
Plasmid classification: Escherichia coli vector; PGEX series expression plasmid
Plasmid size: 5005bp
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
pGEX-KG Plasmid Description
The size of the pGEX-KG carrier is 5005bp, which can grow on the Amp plate, and the GST label can be used to purify the recombinant protein with the GST affinity column
pGEX-KG Plasmid Sequence
LOCUS Exported File 5005 bp ds-DNA circular SYN 29-1-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pGEX-KG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5005)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-1-29
FEATURES Location/Qualifiers
source 1..5005
/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 1308..1412
/gene="bla"
/note="AmpR promoter"
CDS 1413..2273
/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 2444..3032
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3276..3353
/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 3354..4436
/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 4449..4470
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4485..4515
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4523..4539
/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 4547..4563
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4579..4595)
/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 gatccccggg aatttccggt ggtggtggtg
961 gaattctaga ctccatgggt cgactcgagc tcaagcttat tcatcgtgac tgactgacga
1021 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
1081 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
1141 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
1201 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
1261 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
1321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
1381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
1441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
1501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
1561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
1621 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
1681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
1741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
1801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
1861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
1921 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
1981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
2041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
2101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
2161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
2221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
2281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
2341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
2401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
2461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
2521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
2581 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
2641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
2701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
2761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
2821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
2881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
2941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
3001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
3061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
3121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
3181 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
3241 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct
3301 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
3361 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
3421 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
3481 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
3541 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
3601 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
3661 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
3721 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
3781 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
3841 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
3901 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
3961 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
4021 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
4081 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
4141 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
4201 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
4261 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
4321 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
4381 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
4441 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
4501 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
4561 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
4621 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
4681 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
4741 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
4801 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
4861 caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
4921 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
4981 aattattttt gatggcgttg gaatt
//
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