质粒序列
LOCUS Exported 4543 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-82L-Kan.
ACCESSION .
VERSION .
KEYWORDS pQE-81L-Kan
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4543)
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..4543
/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 147..194
/note="MCS"
/note="multiple cloning site"
misc_feature 194..204
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 210..304
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 348..1007
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1072..1158
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1251..2333)
/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(2334..2411)
/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(2929..3517)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3624..4439)
/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 ccatacggat ccgcatgcga gctcggtacc ccgggtcgac
181 ctgcagccaa gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc
241 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
301 gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
361 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
421 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
481 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
541 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
601 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
661 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
721 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
781 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
841 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
901 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
961 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
1021 gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa
1081 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
1141 ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa gctgtcaaac
1201 atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
1261 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
1321 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
1381 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
1441 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
1501 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
1561 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
1621 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
1681 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
1741 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
1801 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
1861 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
1921 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
1981 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
2041 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
2101 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
2161 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
2221 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
2281 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
2341 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat
2401 tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga
2461 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
2521 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
2581 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
2641 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
2701 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
2761 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
2821 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
2881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
3001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
3061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
3121 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
3181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
3241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
3301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
3361 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
3421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
3481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
3541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tggcgttaag
3601 ggattttggt catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
3661 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
3721 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
3781 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
3841 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
3901 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
3961 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
4021 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
4081 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
4141 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
4201 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
4261 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
4321 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
4381 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
4441 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccatgacatt
4501 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//