质粒序列
LOCUS Exported 4758 bp ds-DNA circular SYN 05-DEC-2013
DEFINITION Vector with a lacIq repressor gene for bacterial expression of
6xHis-tagged proteins that can be processed by TAGZyme.
ACCESSION .
VERSION .
KEYWORDS TAGZyme pQE-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4758)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT For TAGZyme cleavage, a glutamine should be inserted at an uneven
amino acid position, followed by the first amino acid of the
expressed protein.
FEATURES Location/Qualifiers
source 1..4758
/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 121..138
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 139..199
/note="MCS"
/note="multiple cloning site"
misc_feature 199..209
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 215..309
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 353..1012
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1077..1163
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1256..2338)
/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(2339..2416)
/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(2934..3522)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3693..4553)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4554..4658)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
121 catcaccatc accatcacca tatgcatgcg ggagctcatg cggccgcggg tacccacgtg
181 tcgacctgca gccaagctta attagctgag cttggactcc tgttgataga tccagtaatg
241 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
301 ggtgagaatc caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa
361 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
421 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
481 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
541 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
601 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
661 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
721 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
781 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
841 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
901 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
961 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
1021 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa
1081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
1141 acgctctcct gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt
1201 caaacatgag aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
1261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
1321 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
1381 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
1441 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
1501 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
1561 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
1621 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
1681 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
1741 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
1801 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
1861 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
1921 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
1981 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
2041 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
2101 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
2161 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
2221 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
2281 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
2341 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
2401 accattcgat ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat
2461 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
2521 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
2581 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
2641 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg
2701 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
2761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
2821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
3361 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
3421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
3601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
3661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
3721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
3781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
3841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
3901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
3961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
4021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
4081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
4141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
4201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
4261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
4321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
4381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
4441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
4501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
4561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
4621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
4681 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
4741 aggccctttc gtcttcac
//