質(zhì)粒名稱:gRNA_Cloning Vector

目錄編號:41824

存儲實(shí)驗(yàn)室:George Church

用途:一個(gè)空的gRNA表達(dá)載體，用于創(chuàng)建一個(gè)特定序列的基因組。載體骨架基本信息載體名稱:pCR-Blunt II-TOPO載體出品公司:Invitrogen載體抗性:卡那霉素載體類型:哺乳細(xì)胞表達(dá),CRISPR高拷貝/低拷貝:未知生長條件:37 ℃插入基因信息克隆方法:TOPO克隆5' 測序引物T7文獻(xiàn)引用方法材料和方法部分:gRNA_Cloning Vector was a gift from George Church plasmid # 41824)

參考文獻(xiàn)部分:RNA-Guided Human Genome Engineering via Cas9. Mali P, Yang L, Esvelt KM, Aach J, Guell M, Dicarlo JE, Norville JE, Church GM. Science. 2013 Jan 3. 10.1126/science.1232033 PubMed 23287722載體質(zhì)粒圖譜和多克隆位點(diǎn)信息gRNA_Cloning Vector

載體圖譜

載體簡介

載體序列LOCUS       gRNA_Cloning Vector Sequencing Result3915 bp DNASYNDEFINITION  gRNA_Cloning Vector Sequencing ResultACCESSION   KEYWORDS    SOURCE        ORGANISM  other sequences; artificial sequences; vectors.FEATURES             Location/Qualifiers     source          1..3915                     /organism="gRNA_Cloning Vector Sequencing Result"                     /mol_type="other DNA"     promoter        143..172                     /label="lac_promoter"     misc_feature    186..208                     /label="M13_pUC_rev_primer"     promoter        207..225                     /label="M13_reverse_primer"     misc_feature    239..256                     /label="Sp6_primer"     promoter        complement(434..678)                     /label="hU6_promoter"     misc_feature    complement(473..492)                     /label="LKO1_5_primer"     promoter        complement(803..821)                     /label="T7_promoter"     misc_feature    825..968                     /label="lacZ_a"     promoter        complement(828..844)                     /label="M13_forward20_primer"     misc_feature    complement(837..859)                     /label="M13_pUC_fwd_primer"     misc_feature    991..1284                     /label="ccdB"     promoter        1495..1544                     /label="NEOKAN_promoter"     CDS             1633..2427                     /label="ORF frame 1"     gene            1636..2424                     /label="NeoR/KanR"                     /gene="NeoR/KanR"     CDS             complement(1942..2475)                     /label="ORF frame 1"     promoter        2561..2589                     /label="AmpR_promoter"     gene            2634..3005                     /label="bleo"                     /gene="bleo"     misc_feature    2634..3008                     /label="sh_ble"     rep_origin      3130..3749                     /label="pBR322_origin"ORIGIN    1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc   61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc  121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa  181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat  241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca  301 ctagtaacgg ccgccagtgt gctggaattc gccctttaat gccaactttg tacaagaaag  361 ctgggtctag aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc  421 cttattttaa cttaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag  481 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta  541 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat  601 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc  661 ctcttcctgc ccgaccttgg taccggatcc agtcgactga attggttcct ttaaagcctg  721 cttttttgta caaagggcga attctgcaga tatccatcac actggcggcc gctcgagcat  781 gcatctagag ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt  841 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat  901 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag  961 ttgcgcagcc tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat 1021 cgtctgtttg tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc 1081 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg 1141 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc 1201 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt 1261 aacctgatgt tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc 1321 tagatccttt tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc 1381 agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc 1441 agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa 1501 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct 1561 ttctcgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga 1621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 1681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 1741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 1801 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 1861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 1921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 1981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 2041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 2101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 2161 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 2221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 2281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 2341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 2401 tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat 2461 tttctcctta cgcatctgtg cggtatttca caccgcatac aggtggcact tttcggggaa 2521 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 2581 tgagacaata accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca 2641 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct 2701 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc 2761 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc 2821 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt 2881 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg 2941 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc 3001 aggactgaca cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 3061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 3121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 3181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 3241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 3301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 3361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 3421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 3481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 3541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 3601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 3661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 3721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc ttttgctggc 3781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 3841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 3901 gcgaggaagc ggaag//