质粒序列
LOCUS Exported File 4952 bp ds-DNA circular SYN 12-4-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pGEX-3X
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4952)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-4-12 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4952
/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 921..932
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
promoter 1255..1359
/gene="bla"
/note="AmpR promoter"
CDS 1360..2220
/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 2391..2979
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3223..3300
/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 3301..4383
/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 4396..4417
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4432..4462
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4470..4486
/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 4494..4510
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4526..4542)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agcttatcga 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 atcgaaggtc gtgggatccc cgggaattca tcgtgactga
961 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
1021 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
1081 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
1141 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
1201 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
1261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
1321 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
1381 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
1441 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
1501 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
1561 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
1621 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
1681 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
1741 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
1801 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
1861 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
1921 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
1981 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
2041 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
2101 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
2161 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
2221 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
2281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
2341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
2401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
2461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
2521 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
2581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
2641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
2701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
2761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
2821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
2881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
2941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
3001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
3061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
3121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
3181 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggtgc
3241 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
3301 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
3361 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
3421 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
3481 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
3541 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
3601 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
3661 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
3721 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
3781 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
3841 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
3901 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
3961 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
4021 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
4081 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
4141 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
4201 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
4261 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
4321 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
4381 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
4441 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
4501 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
4561 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
4621 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
4681 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
4741 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
4801 ccatctacac caacgtaacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
4861 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
4921 agacgcgaat tatttttgat ggcgttggaa tt
//