质粒序列
LOCUS Exported 4005 bp ds-DNA circular SYN 05-DEC-2013
DEFINITION Bacterial vector for expressing DHFR fusion proteins. Similar to
pQE-17, but with a different reading frame after the DHFR.
ACCESSION .
VERSION .
KEYWORDS pQE-18
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4005)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT Discontinued pQE vector.
FEATURES Location/Qualifiers
source 1..4005
/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 133..690
/codon_start=1
/gene="Mus musculus Dhfr"
/product="mouse dihydrofolate reductase"
/note="DHFR"
/translation="MVRPLNSIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
PGVLSEVQEEKGIKYKFEVYEKK"
CDS 711..728
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 735..745
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 751..845
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 889..1548
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1613..1699
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(2181..2769)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2940..3800)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
LIKHW"
promoter complement(3801..3905)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatccggca tcatggttcg accattgaac tcgatcgtcg ccgtgtccca aaatatgggg
181 attggcaaga acggagacct accctggcct ccgctcagga acgagttcaa gtacttccaa
241 agaatgacca caacctcttc agtggaaggt aaacagaatc tggtgattat gggtaggaaa
301 acctggttct ccattcctga gaagaatcga cctttaaagg acagaattaa tatagttctc
361 agtagagaac tcaaagaacc accacgagga gctcattttc ttgccaaaag tttggatgat
421 gccttaagac ttattgaaca accggaattg gcaagtaaag tagacatggt ttggatagtc
481 ggaggcagtt ctgtttacca ggaagccatg aatcaaccag gccaccttag actctttgtg
541 acaaggatca tgcaggaatt tgaaagtgac acgtttttcc cagaaattga tttggggaaa
601 tataaacttc tcccagaata cccaggcgtc ctctctgagg tccaggagga aaaaggcatc
661 aagtataagt ttgaagtcta cgagaagaaa ggttccagat cttccgatct catcaccatc
721 accatcacta agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct
781 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
841 agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
901 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
961 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
1021 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
1081 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
1141 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
1201 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
1261 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
1321 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
1381 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
1441 atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg
1501 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
1561 agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa
1621 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
1681 tctcctgagt aggacaaatc cgccgctcta gagctgcctc gcgcgtttcg gtgatgacgg
1741 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
1801 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
1861 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
1921 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
1981 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtctgt
2041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
2101 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
2161 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
2221 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
2281 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
2341 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
2401 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
2461 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
2521 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
2581 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
2641 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
2701 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
2761 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
2821 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
2881 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
2941 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
3001 gctgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
3061 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
3121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
3181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
3241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
3301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
3361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
3421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
3481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
3541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
3601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
3661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
3721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
3781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
3841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
3901 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
3961 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcac
//