質粒序列
LOCUS Exported 4542 bp ds-DNA circular SYN 05-DEC-2013
DEFINITION Bacterial lacIq vector with a kanamycin resistance marker for
expressing N-terminally 6xHis-tagged proteins. For other reading
frames, use pQE-80L-Kan or pQE-81L-Kan.
ACCESSION .
VERSION .
KEYWORDS pQE-82L-Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4542)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT Because this vector contains the lacIq gene, the pREP4 plasmid is
not needed.
FEATURES Location/Qualifiers
source 1..4542
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 101..106
/note="ribosome binding site"
CDS 115..117
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 146..193
/note="MCS"
/note="multiple cloning site"
misc_feature 193..203
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 209..303
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 347..1006
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1071..1157
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1250..2332)
/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"
promoter complement(2333..2410)
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
rep_origin complement(2928..3516)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3623..4438)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatctcacc atcaccatca ccatgggatc cgcatgcgag ctcggtaccc cgggtcgacc
181 tgcagccaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
241 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
301 aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
361 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
421 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
481 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
541 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
601 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
661 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
721 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
781 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
841 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
901 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
961 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
1021 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
1081 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
1141 tcctgagtag gacaaatccg ccctctagat tacgtgcagt cgatgataag ctgtcaaaca
1201 tgagaattgt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
1261 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
1321 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
1381 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
1441 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
1501 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
1561 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
1621 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
1681 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
1741 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
1801 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
1861 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
1921 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
1981 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
2041 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
2101 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
2161 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
2221 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
2281 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
2341 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
2401 cgatggtgtc ggaatttcgg gcagcgttgg gtcctggcca cgggtgcgca tgatctagag
2461 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
2521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
2581 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
2641 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
2701 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
2761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
2821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
2881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
2941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
3541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat ggcgttaagg
3601 gattttggtc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
3661 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
3721 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
3781 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
3841 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
3901 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
3961 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
4021 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
4081 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
4141 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
4201 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
4261 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
4321 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
4381 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
4441 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac catgacatta
4501 acctataaaa ataggcgtat cacgaggccc tttcgtcttc ac
//