質(zhì)粒序列
LOCUS Exported 6029 bp ds-DNA circular SYN 11-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6029)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-11 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6029
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(884..904)
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/note="TEV site"
/translation="ENLYFQG"
CDS complement(935..952)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 1003..1027
/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(1028..1046)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1355..1432
/gene="lacI"
/note="lacI promoter"
CDS 1433..2515
/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 3324..3515
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3617..3759
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3945..4533)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4655..5470
/codon_start=1
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5563..6018)
/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 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccggtacca ctagttagag accaagacac gccttgtgac
241 tgtcctgcag ctttattctc ttgatgctgg tgctggaata gccctcatca ctgccgaggc
301 tctgcatgct gccccgctcg tcagagtcgc tcacactgct gctgctccag tccagatcac
361 ctgtgagata gtccgtgctc tccacgtcaa cgtcgatttc ttccctgtcg gagtcggagc
421 gctccgagga gacggtggag ccgatgctgt ccatccggat cctctcaatg cccagcttct
481 ccagctgcct cttcaggtgt cgctgctctc gctgaagctg gtcgatttgg tgaacggctt
541 ttctgtcaca atcttcaagt ttctttatgt gcaatttggc ttttgttaat aaactcaacg
601 tagtgtgtcg acttgattcg ggtcccagtg gcaccagccc cttcaacttc tccaggcaca
661 agcgaagatg agcccgtcta ttcttctcca tttcattgtg agttgatctg ctactgctgt
721 tattcttttt ggatttgttc ctccgtttta aggcatctct gtccttgttt ttgtatggta
781 acatggaggc ataaccatgt tcagcttctc tctcccgccg ctccagatag tcggccgcct
841 ccagcagcat ctggatgttc atccgaaccg ccgccgccat ggcgccctga aaataaagat
901 tctcagtagt ggggatgtcg taatcgctca tggggtgatg gtgatggtga tgtttcatgg
961 tatatctcct tcttaaagtt aaatcaaaat tatttctaga ggggaattgt tatccgctca
1021 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
1081 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
1141 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
1201 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
1261 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
1321 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
1381 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
1441 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
1501 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
1561 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
1621 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
1681 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
1741 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
1801 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
1861 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
1921 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
1981 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
2041 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
2101 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
2161 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
2221 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
2281 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
2341 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
2401 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
2461 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
2521 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
2581 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
2641 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
2701 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
2761 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
2821 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
2881 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
2941 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
3001 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
3061 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
3121 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
3181 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
3241 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
3301 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
3361 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
3421 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
3481 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
3541 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
3601 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
3661 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
3721 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
3781 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
3841 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
3901 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
3961 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4021 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4081 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4141 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4201 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4261 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4321 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4381 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4441 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4501 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4561 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
4621 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
4681 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
4741 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
4801 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
4861 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
4921 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
4981 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
5041 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
5101 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
5161 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
5221 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
5281 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
5341 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
5401 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
5461 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
5521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
5581 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
5641 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
5701 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5761 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5821 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5881 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
5941 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
6001 cgccgctaca gggcgcgtcc cattcgcca
//