质粒序列
LOCUS Exported 5760 bp ds-DNA circular SYN 05-MAY-2014
DEFINITION Vector for expression in bacterial, insect, and vertebrate cells of
proteins with C-terminal Strep-Tag II and 8xHis tags.
ACCESSION .
VERSION .
KEYWORDS pQE-TriSystem His-Strep1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5760)
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..5760
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 62..367
/note="CMV enhancer"
/note="human cytomegalovirus (CMV) immediate early
enhancer"
promoter 373..650
/note="chicken beta-actin promoter"
exon 651..742
/note="Exon 1"
intron 743..1004
exon 1005..1613
/note="Exon 2"
promoter 1073..1116
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 1092..1108
/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 1143..1252
/note="p10 promoter"
/note="baculovirus promoter for expression in insect cells"
RBS 1255..1260
/note="ribosome binding site"
misc_feature 1265..1372
/note="MCS"
/note="multiple cloning site"
CDS 1267..1269
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 1381..1404
/codon_start=1
/product="peptide that binds Strep-Tactin, an engineered
form of streptavidin"
/note="Strep-Tag II"
/translation="WSHPQFEK"
CDS 1411..1434
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
misc_feature 1435..1445
/note="stop codons"
/note="stop codons in all three reading frames"
polyA_signal 1589..1594
terminator 1611..1722
/note="rabbit globin terminator"
terminator 1732..1779
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 1792..3250
/note="baculovirus recombination region (ORF1629)"
/note="contains part of ORF1629"
CDS complement(1909..3250)
/codon_start=2
/product="baculovirus capsid-associated protein"
/note="ORF1629"
/note="required for viral replication"
/translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
rep_origin complement(3409..3996)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4009..4869)
/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"
misc_recomb 4935..2
/note="baculovirus recombination region (lef2/ORF603)"
/note="contains ORF603 and part of lef2"
CDS 4935..5059
/codon_start=3
/product="baculovirus late expression factor 2"
/note="lef2"
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(5097..5702)
/codon_start=1
/product="baculovirus ORF603 protein"
/note="ORF603"
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
1 ctctagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
361 taccatgcat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
421 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
481 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
541 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
601 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgtgc
661 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
721 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctt cgggctgtaa
781 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
841 gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct
901 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
961 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
1021 tggttattgt gctgtctcat cattttggca aagaattgga tcggaccgga aatcataaaa
1081 aattatttgc tttgtgagcg gataacaatt ataatagatt caattgaggc ctcgaccacc
1141 gggaccttta attcaaccca acacaatata ttatagttaa ataagaatta ttatcaaatc
1201 atttgtatat taattaaaat actatactgt aaattacatt ttatttacaa tcaaaggaga
1261 tataccatgg cgatatcccg ggagctcgtg gatccgaatt ctcagatctc ggcgcgcctg
1321 caggtcgacg gtaccggttc gaagcttgcg gccgcacagc tgtatacacg tgcaagcgcg
1381 tggagccacc cgcagttcga aaaactcgag caccaccatc accatcacca tcactaagtg
1441 attaacctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
1501 ctcacaaata ccactgagat cgatcttttt ccctctgcca aaaattatgg ggacatcatg
1561 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
1621 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
1681 catcagaatg agtttttggt ttagagtttg gcaacatatg cccatatgta actagcataa
1741 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagcatg cggaggaaat
1801 tctccttgaa gtttccctgg tgttcaaagt aaaggagttt gcaccagacg cacctctgtt
1861 cactggtccg gcgtattaaa acacgataca ttgttattag tacatttatt aagcgctaga
1921 ttctgtgcgt tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt
1981 ataatcttta gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct
2041 gaatttaaat attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa
2101 gggttgtttt tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa
2161 acttgccaaa tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg
2221 taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc
2281 gttagtgtac aattgactcg acgtaaacac gttaaataga gcttggacat atttaacatc
2341 gggcgtgtta gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt
2401 tgcttccgaa gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac
2461 gtccgcgatc aaatttgtag ttgagctttt tggaattatt tctgattgcg ggcgtttttg
2521 ggcgggtttc aatctaactg tgcccgattt taattcagac aacacgttag aaagcgatgg
2581 tgcaggcggt ggtaacattt cagacggcaa atctactaat ggcggcggtg gtggagctga
2641 tgataaatct accatcggtg gaggcgcagg cggggctggc ggcggaggcg gaggcggagg
2701 tggtggcggt gatgcagacg gcggtttagg ctcaaatgtc tctttaggca acacagtcgg
2761 cacctcaact attgtactgg tttcgggcgc cgtttttggt ttgaccggtc tgagacgagt
2821 gcgatttttt tcgtttctaa tagcttccaa caattgttgt ctgtcgtcta aaggtgcagc
2881 gggttgaggt tccgtcggca ttggtggagc gggcggcaat tcagacatcg atggtggtgg
2941 tggtggtgga ggcgctggaa tgttaggcac gggagaaggt ggtggcggcg gtgccgccgg
3001 tataatttgt tctggtttag tttgttcgcg cacgattgtg ggcaccggcg caggcgccgc
3061 tggctgcaca acggaaggtc gtctgcttcg aggcagcgct tggggtggtg gcaattcaat
3121 attataattg gaatacaaat cgtaaaaatc tgctataagc attgtaattt cgctatcgtt
3181 taccgtgccg atatttaaca accgctcaat gtaagcaatt gtattgtaaa gagattgtct
3241 caagctcgga acgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
3301 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
3361 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
3421 ccgccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgtt accaatgctt
4021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
4081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
4141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
4201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
4261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
4321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
4381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
4441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
4501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
4561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
4621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
4681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
4741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
4801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
4861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
4921 gtccgcgcgt ttcctgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg
4981 ccaaaaaatg aaaactgtcg acaagctctg tccgtttgct ggcaactgca agggtctcaa
5041 tcctatttgt aattattgaa taataaaaca attataaatg tcaaatttgt tttttattaa
5101 cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag
5161 ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac
5221 aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc
5281 tctttttcat ttttctcttc ataaaaatta acatagttat tatcgtatcc atatatgtat
5341 ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt
5401 atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct
5461 tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt
5521 ttgtacaata tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt
5581 agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt
5641 tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc
5701 attgtaatga gacgcacaaa ctaatatcac aaactggaaa tgtctatcaa tatatagttg
//