质粒序列
LOCUS Exported 5308 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-24c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5308)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5308
/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(205..237)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
protein_bind 282..306
/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(307..325)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 634..711
/gene="lacI"
/note="lacI promoter"
CDS 712..1794
/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 2603..2794
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3224..3812)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS 3934..4749
/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(4842..5297)
/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 tccgacccat ttgctgtcca ccagtcatgc tagccatatg
241 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
301 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg
361 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
421 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
481 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
541 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
601 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
661 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
721 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
781 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
841 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
901 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
961 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
1021 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
1081 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
1141 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
1201 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
1261 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
1321 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
1381 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
1441 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
1501 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
1561 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
1621 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
1681 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
1741 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
1801 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag
1861 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
1921 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
1981 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
2041 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
2101 cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct
2161 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
2221 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
2281 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
2341 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
2401 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
2461 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
2521 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
2581 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
2641 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
2701 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
2761 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
2821 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
2881 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
2941 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa
3001 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
3661 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
3841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
3901 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
3961 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
4021 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
4081 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
4141 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
4201 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
4261 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
4321 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
4381 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
4441 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
4501 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
4561 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
4621 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
4681 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
4741 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca
4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat
4861 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
4921 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
4981 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
5041 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
5101 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
5161 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
5221 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
5281 gccgctacag ggcgcgtccc attcgcca
//