• 规格：20μl质粒
• 启动子：hH1 promoter
• 原核抗性：卡那霉素Kan
• 筛选标记：遗传霉素G418
• 克隆菌株：大肠杆菌DH5α
• 质粒简介：           phH1-gRNA是一个哺乳细胞gRNA表达基因编辑质粒。
• 质粒图谱1： png/1-1P10G11043560.png

质粒序列： 点击查看序列

LOCUS       Exported                3451 bp ds-DNA     circular SYN 07-JAN-2018

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..3451

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        1..215

                     /label=H1 promoter

                     /note="human H1 RNA promoter"

     misc_RNA        242..317

                     /label=gRNA scaffold

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

     rep_origin      complement(803..1391)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(1468..2262)

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /label=NeoR/KanR

                     /note="confers resistance to neomycin, kanamycin, and G418 

                     (Geneticin(R))"

                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     promoter        complement(2297..2654)

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2311..2446

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     promoter        complement(2656..2760)

                     /gene="bla"

                     /label=AmpR promoter

     rep_origin      complement(2786..3241)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

ORIGIN

        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa

       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc

      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg

      181 gatttgggaa tcttataagt tctgtatgag accactcttt cccaggtctt cgagaagacc

      241 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag

      301 tggcaccgag tcggtgcttt ttttggtctg agacgccgcg gtggagctcc agcttttgtt

      361 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt

      421 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag

      481 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt

      541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag

      601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg

      661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat

      721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta

      781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa

      841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc

      901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt

      961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca

     1021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg

     1081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat

     1141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta

     1201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct

     1261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac

     1321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa

     1381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa

     1441 actcacgtta agggattttg gtcatgatca gaagaactcg tcaagaaggc gatagaaggc

     1501 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc

     1561 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc

     1621 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt

     1681 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt

     1741 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg

     1801 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg

     1861 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat

     1921 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc

     1981 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac

     2041 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc

     2101 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc

     2161 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca

     2221 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc

     2281 tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct

     2341 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc

     2401 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg

     2461 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg

     2521 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc

     2581 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt

     2641 tctttccgcc tcaggactct tcctttttca atattattga agcatttatc agggttattg

     2701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg

     2761 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg

     2821 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct

     2881 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt

     2941 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat

     3001 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca

     3061 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac

     3121 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta

     3181 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg

     3241 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg

     3301 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca

     3361 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca

     3421 ctatagggcg aattgggtac ccgtctcacc t

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