質(zhì)粒序列
LOCUS Exported 3512 bp ds-DNA circular SYN 05-DEC-2013
DEFINITION Bacterial vector for expressing proteins with an N-terminal 6xHis
tag and a C-terminal Tag-100 tag.
ACCESSION .
VERSION .
KEYWORDS pQE-100 DoubleTag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3512)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..3512
/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 145..184
/note="MCS"
/note="multiple cloning site"
CDS 193..228
/codon_start=1
/product="epitope tag derived from MAP kinase 2"
/note="Tag-100"
/translation="EETARFQPGYRS"
misc_feature 243..253
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 259..353
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 397..1056
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1121..1207
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(1688..2276)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2447..3307)
/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(3308..3412)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatcgcatc accatcacca tcacggatcc gcatgcgagc tcggtacccc gggtcgacct
181 gcagccaagc ttgaagagac tgcacgtttc cagccgggtt atcgttctta gagatctaag
241 cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
301 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct
361 agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
421 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
481 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
541 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
601 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
661 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
721 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
781 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
841 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
901 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
961 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
1021 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
1081 ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac gaaaggctca
1141 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
1201 gacaaatccg ccctctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
1261 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
1321 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
1381 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
1441 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
1501 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
1561 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
1621 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
1681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
1741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
1801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
1861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
1921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
1981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
2041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
2101 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
2161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
2221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
2281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
2341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
2401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
2461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
2521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
2581 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
2641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
2701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
2761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
2821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
2881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
2941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
3001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
3061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
3121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
3181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
3241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
3301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
3361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
3421 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
3481 ataggcgtat cacgaggccc tttcgtcttc ac
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