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基本信息

質粒圖譜和多克隆位點信息 

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載體描述

pQE-30 pQE vectors and constructs can be maintained in any E. coli strain that is ampicillin-sensitive and carries the pREP4 repressor plasmid, or harbors the lacIq gene on the F-factor episome.

載體序列

LOCUS       Exported                3461 bp ds-DNA     circular SYN 16-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 3461)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, Mar 16, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..3461

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        10..54

                     /label=T5 promoter

                     /note="bacteriophage T5 promoter for E. coli RNA

                     polymerase, with embedded lac operator"

     protein_bind    30..46

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /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

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /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)."

     primer_bind     67..88

                     /label=pQE30-F

     RBS             97..108

                     /note="strong bacterial ribosome binding site (Elowitz and

                     Leibler, 2000)"

     CDS             127..144

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     misc_feature    145..192

                     /label=MCS

     terminator      208..302

                     /label=lambda t0 terminator

                     /note="transcription terminator from phage lambda"

     primer_bind     complement(222..242)

                     /label=pQE30-R

     CDS             346..1005

                     /codon_start=1

                     /gene="cat"

                     /product="chloramphenicol acetyltransferase"

                     /label=CmR

                     /note="confers resistance to chloramphenicol"

                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL

                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS

                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM

                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"

     terminator      1070..1156

                     /gene="Escherichia coli rrnB"

                     /label=rrnB T1 terminator

                     /note="transcription terminator T1 from the E. coli rrnB

                     gene"

     misc_feature    1311..1451

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(1637..2225)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

     CDS             complement(2396..3256)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(3257..3361)

                     /gene="bla"

                     /label=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 ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag

      241 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga

      301 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc

      361 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt

      421 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta

      481 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc

      541 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg

      601 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc

      661 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg

      721 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc

      781 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac

      841 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg

      901 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt

      961 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt

     1021 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg

     1081 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct

     1141 cctgagtagg acaaatccgc cctctagagc tgcctcgcgc gtttcggtga tgacggtgaa

     1201 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg

     1261 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg

     1321 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga

     1381 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat

     1441 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc

     1501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg

     1561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg

     1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac

     1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg

     1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct

     1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg

     1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct

     1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac

     1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt

     2041 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc

     2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca

     2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat

     2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac

     2281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt

     2341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc

     2401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg

     2461 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg

     2521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc

     2581 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta

     2641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg

     2701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct

     2761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta

     2821 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg

     2881 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga

     2941 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt

     3001 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca

     3061 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt

     3121 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt

     3181 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga

     3241 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt

     3301 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc

     3361 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa

     3421 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c

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