• 规格：20μl质粒
• 复制子：pUC
• 原核抗性：卡那霉素Kan
• 筛选标记：致死基因ccdB
• 克隆菌株：大肠杆菌DB3.1
• 质粒简介：             pENTR1A Dual是一个Gateway系统入门克隆质粒。
• 质粒图谱1： png/1-1G214160I45K.png

质粒序列： 点击查看序列

LOCUS       Exported                3754 bp ds-DNA     circular SYN 14-DEC-2017

DEFINITION  Gateway(R) Dual Selection Vector for creating an entry vector by 

            restriction cloning. Other reading frames are available with 

            pENTR(TM)2B and pENTR(TM)3C.

COMMENT     To create a Kozak sequence, clone a blunt fragment starting with 

            ATGG (where ATG is the start codon) into the XmnI site.

FEATURES             Location/Qualifiers

     source          1..3754

                     /organism="synthetic DNA construct"

                     /lab_host="Escherichia coli"

                     /mol_type="other DNA"

     terminator      103..189

                     /gene="Escherichia coli rrnB"

                     /label=rrnB T1 terminator

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

                     gene"

     terminator      281..308

                     /label=rrnB T2 terminator

                     /note="transcription terminator T2 from the E. coli rrnB 

                     gene"

     protein_bind    358..457

                     /gene="mutant version of attL"

                     /label=attL1

                     /bound_moiety="LR Clonase(TM)"

                     /note="recombination site for the Gateway(R) LR reaction"

     misc_feature    457..507

                     /label=MCS 1

                     /note="multiple cloning site, part 1"

     RBS             461..466

                     /note="ribosome binding site"

     promoter        524..554

                     /label=lac UV5 promoter

                     /note="E. coli lac promoter with an ""up"" mutation"

     CDS             608..1288

                     /codon_start=1

                     /product="chloramphenicol acetyltransferase"

                     /label=CmR

                     /note="confers resistance to chloramphenicol"

                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL

                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS

                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM

                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE

                     DPAY"

     CDS             1608..1913

                     /codon_start=1

                     /gene="ccdB"

                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"

                     /label=ccdB

                     /note="Plasmids containing the ccdB gene cannot be 

                     propagated in standard E. coli strains."

                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK

                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"

     misc_feature    1948..1979

                     /label=MCS 2

                     /note="multiple cloning site, part 2"

     protein_bind    complement(1983..2082)

                     /gene="mutant version of attL"

                     /label=attL2

                     /bound_moiety="LR Clonase(TM)"

                     /note="recombination site for the Gateway(R) LR reaction"

     CDS             2205..3014

                     /codon_start=1

                     /gene="aph(3')-Ia"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin in bacteria or G418 

                     (Geneticin(R)) in eukaryotes"

                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP

                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA

                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD

                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD

                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     rep_origin      3104..3692

                     /direction=RIGHT

                     /label=ori

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

                     replication"

ORIGIN

        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg

       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct

      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt

      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg

      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat

      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa

      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc

      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac

      481 tggatccggt accgaattcg cggccgcatt aggcacccca ggctttacac tttatgcttc

      541 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga

      601 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg

      661 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca

      721 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc

      781 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa

      841 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca

      901 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca

      961 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt

     1021 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt

     1081 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac

     1141 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg

     1201 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg

     1261 gcgtaatcta gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct

     1321 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt

     1381 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg

     1441 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg

     1501 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt

     1561 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg

     1621 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta

     1681 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata

     1741 aagtcccccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga

     1801 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc

     1861 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag

     1921 gctcccttat acacagccag tctgcaggtc gacgaattcg cggccgcact cgagatatct

     1981 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg

     2041 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct

     2101 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa

     2161 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac

     2221 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat

     2281 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg

     2341 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg

     2401 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta

     2461 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc

     2521 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgtccc

     2581 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc

     2641 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg

     2701 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat

     2761 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg

     2821 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg

     2881 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt

     2941 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg

     3001 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat tgggccccgt

     3061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc

     3121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc

     3181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac

     3241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac

     3301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt

     3361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct

     3421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat

     3481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt

     3541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg

     3601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt

     3661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt

     3721 tcctggcctt ttgctggcct tttgctcaca tgtt

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