质粒序列
LOCUS Exported 5931 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-42c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5931)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5931
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(150..173)
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
CDS complement(266..277)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
CDS complement(308..352)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(368..385)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(395..412)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(440..1093)
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
protein_bind 1138..1162
/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)."
promoter complement(1163..1181)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1494..1571
/gene="lacI"
/note="lacI promoter"
CDS 1572..2654
/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"
CDS 3226..3417
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3847..4435)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS 4557..5372
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5465..5920)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
241 ggatccccga tatcgccatg gacccgcgtc cctcaatacc ggagccacca ccggtaccca
301 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccaa
361 ttgcagtact accgcgtggc accagacccg cggagtgatg gtgatggtga tgaccagaac
421 cactagttga accatccgat tttggaggat ggtcgccacc accaaacgtg gcttgccagc
481 cctgcaaagg ccatgctata tacttgctgg atttcaagta cttatcaatt tgtgggatag
541 cttcaatacg ttttttaaaa caaactaatt ttgggaacgc atccaggcac attgggtcca
601 tgtataaaac aacatcaaga gcgtcataca acatgaagtc aggatgggtt acatgatcac
661 catttaaata tgttttatga cataaacgat cttcgaacat tttcagcatt tcaggtagct
721 tgctaagaaa atcaactttg agagtttcaa agtctttact atatgcaatt ctcgaaacac
781 cgtatctaat atccaaaacc gctccttcaa gcattgaaat ctctgcacgc tcttttggac
841 aaccacccaa catgttgtgc ttgtcagcta tataacgtat gatggccata gactgtgtta
901 atttaacatc accatcaata taataaggaa gattgggaaa ctccaaaccc aattcaaact
961 ttttgtttcg ccatttatca ccttcatcgc gctcatacaa atgctcttca tatttttctt
1021 caagatattc caaaagaagt cgagtgggtt gcacaaggcc cttaattttc caataaccta
1081 gtatagggga catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
1141 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
1201 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
1261 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
1321 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
1381 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
1441 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
1501 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
1561 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
1621 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
1681 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
1741 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
1801 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
1861 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
1921 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
1981 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
2041 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
2101 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
2161 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
2221 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
2281 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
2341 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
2401 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
2461 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
2521 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
2581 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
2641 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
2701 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
2761 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
2821 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
2881 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
2941 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
3001 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
3061 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
3121 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
3181 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
3241 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
3301 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
3361 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
3421 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
3481 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
3541 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
3601 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
3661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
3721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
3781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
3841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
3901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
3961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
4021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
4081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
4141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
4201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
4261 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
4321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
4381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
4441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
4501 tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
4561 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
4621 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
4681 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
4741 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
4801 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
4861 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
4921 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
4981 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
5041 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
5101 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
5161 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
5221 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
5281 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
5341 agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg atacatattt
5401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
5461 cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
5521 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
5581 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
5641 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
5701 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
5761 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
5821 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
5881 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a
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