質(zhì)粒序列
LOCUS Exported 5302 bp ds-DNA circular SYN 05-MAY-2014
DEFINITION Vector for expressing C-terminally 10xHis-tagged proteins in insect
and mammalian cells using the QIAgenes system.
ACCESSION .
VERSION .
KEYWORDS pQE-Trisystem-6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5302)
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..5302
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_recomb 181..1008
/note="baculovirus recombination region (lef2/ORF603)"
/note="contains ORF603 and part of lef2"
CDS 181..305
/codon_start=3
/product="baculovirus late expression factor 2"
/note="lef2"
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(343..948)
/codon_start=1
/product="baculovirus ORF603 protein"
/note="ORF603"
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
enhancer 1138..1443
/note="CMV enhancer"
/note="human cytomegalovirus (CMV) immediate early
enhancer"
promoter 1449..1726
/note="chicken beta-actin promoter"
exon 1727..1819
/note="Exon 1"
intron 1820..2081
exon 2082..2686
/note="Exon 2"
promoter 2150..2168
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2169..2193
/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 2209..2318
/note="p10 promoter"
/note="baculovirus promoter for expression in insect cells"
RBS 2321..2326
/note="ribosome binding site"
CDS 2338..2340
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
misc_feature 2347..2404
/note="MCS"
/note="multiple cloning site"
CDS 2415..2444
/codon_start=1
/product="10xHis affinity tag"
/note="10xHis"
/translation="HHHHHHHHHH"
misc_feature 2445..2450
/note="stop codons"
/note="two tandem stop codons"
misc_feature 2459..2477
/note="siRNA target site"
CDS 2484..2507
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
misc_feature 2508..2518
/note="stop codons"
/note="stop codons in all three reading frames"
polyA_signal 2662..2667
terminator 2684..2795
/note="rabbit globin terminator"
terminator 2805..2852
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_feature 2865..3570
/note="baculovirus recombination region (ORF1629)"
/note="contains part of ORF1629"
CDS complement(2982..3570)
/codon_start=2
/product="baculovirus capsid-associated protein"
/note="ORF1629"
/note="required for viral replication"
/translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
rep_origin complement(3780..4367)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4380..5240)
/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"
ORIGIN
1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccg
1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct
1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
1561 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
1621 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
1681 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
1741 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
1801 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctccttcgg gctgtaatta
1861 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
1921 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc
1981 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
2041 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
2101 ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt aatacgactc
2161 actatagggg aattgtgagc ggataacaat tccccggagt taatccggga cctttaattc
2221 aacccaacac aatatattat agttaaataa gaattattat caaatcattt gtatattaat
2281 taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata ccatgatatg
2341 caggtcgata tcccgggtgg atccgaattc tcagatctcg gcgcgcctgc agggttcgaa
2401 gcttacctgc tatacaccac caccatcacc atcatcacca ccactagtga ctcgagaagc
2461 gttgaaatag cgtacaagtc gagcaccacc atcaccatca ccatcactaa gtgattaacc
2521 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
2581 ataccactga gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
2641 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
2701 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
2761 atgagtattt ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg
2821 gggcctctaa acgggtcttg aggggttttt tgctgaaagc atgcggagga aattctcctt
2881 gaagtttccc tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt
2941 ccggcgtatt aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg
3001 cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct
3061 ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta
3121 aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt
3181 ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc
3241 aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa
3301 ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg
3361 tacaattgac tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg
3421 ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc
3481 gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg
3541 atcaaatttg tagttgagct ttttggaatt gcgatcgcat aacttcgtat agcatacatt
3601 atacgaagtt ataagctcgg aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca
3661 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
3721 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
3781 ttccataggc tccgccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3961 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttgt
4381 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
4441 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
4501 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
4561 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
4621 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
4681 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
4741 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
4801 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
4861 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
4921 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
4981 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5041 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5101 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5161 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
5221 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
5281 tattgtctca tgtccgcgcg tt
//