質(zhì)粒序列
LOCUS Exported 5317 bp ds-DNA circular SYN 09-JAN-2013
DEFINITION Vector for expressing N-terminally 10xHis-tagged proteins in
bacteria using the QIAgenes system.
ACCESSION .
VERSION .
KEYWORDS pQE-T7-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5317)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT A QIAgene can be cloned into this vector as an NdeI-XhoI fragment.
FEATURES Location/Qualifiers
source 1..5317
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/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"
misc_feature 158..217
/note="MCS"
/note="multiple cloning site"
misc_feature 166..171
/note="stop codons"
/note="two tandem stop codons"
CDS complement(215..244)
/codon_start=1
/product="10xHis affinity tag"
/note="10xHis"
/translation="HHHHHHHHHH"
CDS complement(248..250)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 258..263
/note="ribosome binding site"
protein_bind 295..319
/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(320..338)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 647..724
/gene="lacI"
/note="lacI promoter"
CDS 725..1807
/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 2616..2807
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3234..3822)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3944..4759
/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(4851..5306)
/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 gagtcttact aggctgcaga
181 cgtgggtacc cgcggccgca tgagctccat gcatatgatg atggtggtga tggtgatgat
241 gatgtttcat atttatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt
301 gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg
361 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
421 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
481 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
541 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
601 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg
661 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
721 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
781 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
841 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
901 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
961 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
1021 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
1081 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
1141 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
1201 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
1261 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
1321 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
1381 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
1441 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
1501 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
1561 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
1621 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
1681 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
1741 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
1801 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac
1861 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
1921 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
1981 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
2041 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
2101 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca
2161 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
2221 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
2281 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
2341 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
2401 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
2461 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
2521 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
2581 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
2641 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
2701 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
2761 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
2821 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
2881 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
2941 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
3001 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
3061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
3121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
3181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
3241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
3301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
3361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
3421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
3481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
3541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
3601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
3661 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
3721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
3781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
3841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac
3901 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
3961 ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
4021 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
4081 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
4141 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
4201 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
4261 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
4321 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
4381 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
4441 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
4501 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
4561 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
4621 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
4681 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
4741 gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga
4801 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
4861 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
4921 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
4981 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
5041 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
5101 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
5161 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
5221 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
5281 gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//