質(zhì)粒序列
LOCUS Exported 3996 bp ds-DNA circular SYN 05-DEC-2013
DEFINITION Bacterial vector for expressing DHFR fusion proteins with a
C-terminal 6xHis tag. Similar to pQE-16 and pQE-22, but with a
different reading frame after the DHFR.
ACCESSION .
VERSION .
KEYWORDS pQE-19
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3996)
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..3996
/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 702..719
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 726..736
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 742..836
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 880..1539
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1604..1690
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(2172..2760)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2931..3791)
/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(3792..3896)
/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 ggttcagatc tcatcaccat caccatcact
721 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
781 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
841 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
901 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
961 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
1021 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
1081 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
1141 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
1201 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
1261 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
1321 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
1381 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
1441 gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa
1501 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
1561 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
1621 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
1681 taggacaaat ccgccgctct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
1741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
1801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
1861 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
1921 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
1981 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtctg tcggctgcgg
2041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
2101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
2161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
2221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
2281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
2341 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
2401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
2461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
2521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
2581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
2641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
2701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
2761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
2821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
2881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
2941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agctgcctga
3001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
3061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
3121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
3181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
3241 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
3301 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
3361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
3421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
3481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
3541 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
3601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
3661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
3721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
3781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
3841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
3901 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
3961 aaaaataggc gtatcacgag gccctttcgt cttcac
//