• 规格：20μl质粒
• 启动子：T7 promoter
• 复制子：pUC ori
• 质粒大小：2792bp
• 原核抗性：卡那霉素Kan
• 克隆菌株：大肠杆菌DH5α
• 质粒简介：         pUC57-sgRNA质粒是一个用于体外转录sgRNA的载体，gRNA片段可通过BsaI酶切连入，并在T7启动子的驱动下转录出来。本质粒的参考文献是：Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects. 
• 质粒图谱1： png/1-1G026153934645.png

质粒序列： 点击查看序列

LOCUS       Exported                2792 bp ds-DNA     circular SYN 26-OCT-2017

DEFINITION  synthetic circular DNA

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 2792)

  AUTHORS   .

  TITLE     Direct Submission

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..2792

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        442..460

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_RNA        482..557

                     /label=gRNA scaffold

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

     rep_origin      complement(1018..1606)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(1777..2592)

                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG

                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK

                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA

                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI

                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     promoter        complement(2593..2697)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc

      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc

      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat

      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt

      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa

      421 tgcatctaga tatcggatcc ctaatacgac tcactatagg tgagaccgag agagggtctc

      481 agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag

      541 tggcaccgag tcggtgcttt ttttaaaggg cccgtcgact gcagaggcct gcatgcaagc

      601 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca

      661 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa

      721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag

      781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgcggccgc

      841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc

      901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat

      961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt

     1021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg

     1081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc

     1141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt

     1201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa

     1261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta

     1321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa

     1381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa

     1441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt

     1501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt

     1561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat

     1621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat

     1681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc

     1741 aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa

     1801 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt

     1861 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg

     1921 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat

     1981 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag

     2041 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc

     2101 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg

     2161 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc

     2221 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt

     2281 tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt

     2341 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac

     2401 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc

     2461 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc

     2521 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg

     2581 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct

     2641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac

     2701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta

     2761 taaaaatagg cgtatcacga ggccctttcg tc

//