質(zhì)粒序列
LOCUS Exported 5291 bp ds-DNA circular SYN 09-JAN-2013
DEFINITION Vector for expressing C-terminally 10xHis-tagged proteins in
bacteria using the QIAgenes system.
ACCESSION .
VERSION .
KEYWORDS pQE-T7-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5291)
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..5291
/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"
misc_feature 137..142
/note="stop codons"
/note="two tandem stop codons"
CDS complement(143..172)
/codon_start=1
/product="10xHis affinity tag"
/note="10xHis"
/translation="HHHHHHHHHH"
misc_feature 173..224
/note="MCS"
/note="multiple cloning site"
CDS complement(219..221)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 232..237
/note="ribosome binding site"
protein_bind 269..293
/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(294..312)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 621..698
/gene="lacI"
/note="lacI promoter"
CDS 699..1781
/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 2590..2781
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3208..3796)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3918..4733
/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(4825..5280)
/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 cggatcttac taatgatgat ggtggtgatg gtgatgatga tgctcgaggg
181 ctgcagacgt gggtacccgc ggccgcatga gctccatgca tatgatttat atctccttct
241 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag
301 tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc
361 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
421 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
481 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
541 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
601 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
661 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
721 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
781 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
841 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
901 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
961 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
1021 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
1081 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
1141 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
1201 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
1261 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
1321 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
1381 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
1441 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
1501 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
1561 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
1621 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
1681 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
1741 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
1801 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
1861 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
1921 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
1981 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
2041 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
2101 attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
2161 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
2221 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
2281 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
2341 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
2401 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
2461 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
2521 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
2581 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
2641 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
2701 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
2761 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
2821 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
2881 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
2941 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
3001 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
3061 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
3121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
3181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
3241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
3301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
3361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
3421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
3481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
3541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
3601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
3661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
3721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
3781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
3841 cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct tacataaaca
3901 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct aggccgcgat
3961 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
4021 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
4081 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
4141 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
4201 ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg
4261 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
4321 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
4381 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
4441 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
4501 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
4561 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
4621 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
4681 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taagaattaa
4741 ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
4801 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
4861 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
4921 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
4981 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
5041 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
5101 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
5161 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
5221 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
5281 cccattcgcc a
//