• 规格：20μl质粒
• 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：          phU6-gRNA是一个哺乳细胞gRNA表达质粒。
• 质粒图谱1： png/1-1G214164636453.png

质粒序列： 点击查看序列

LOCUS       Exported                3515 bp ds-DNA     circular SYN 30-NOV-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..3515

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      complement(3..458)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        696..936

                     /label=U6 promoter

                     /note="RNA polymerase III promoter for human U6 snRNA"

     misc_RNA        963..1038

                     /label=gRNA scaffold

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

     rep_origin      complement(1535..2123)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(2200..2994)

                     /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(3029..3386)

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      3043..3178

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     promoter        complement(3388..3492)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc

       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga

      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc

      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc

      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag

      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa

      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac

      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg

      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg

      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg

      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacccg

      661 tctcacaggc ggatcgatcc aaggtcgggc aggaagaggg cctatttccc atgattcctt

      721 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa

      781 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg

      841 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt

      901 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgggtct tcgagaagac

      961 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa

     1021 gtggcaccga gtcggtgctt tttttctccg ctgagcgtac tgagacgccg cggtggagct

     1081 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc

     1141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca

     1201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct

     1261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac

     1321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc

     1381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt

     1441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg

     1501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg

     1561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat

     1621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta

     1681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct

     1741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc

     1801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa

     1861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg

     1921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag

     1981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt

     2041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta

     2101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc

     2161 agtggaacga aaactcacgt taagggattt tggtcatgat cagaagaact cgtcaagaag

     2221 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg

     2281 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg

     2341 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc

     2401 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg

     2461 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc

     2521 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg

     2581 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc

     2641 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc

     2701 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc

     2761 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc

     2821 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag

     2881 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag

     2941 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa

     3001 cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc

     3061 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa

     3121 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg

     3181 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc

     3241 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc

     3301 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat

     3361 tccacagctg gttctttccg cctcaggact cttccttttt caatattatt gaagcattta

     3421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat

     3481 aggggttccg cgcacatttc cccgaaaagt gccac

//