gRNA_Cloning Vector

gRNA_Cloning Vector

载体基本信息

质粒名称:	gRNA_Cloning Vector
出品公司:	Ybscience
存储实验室:	George Church
用途:	一个空的gRNA表达载体，用于创建一个特定序列的基因组。
载体骨架基本信息
载体名称:	pCR-Blunt II-TOPO
载体出品公司:	Invitrogen
载体抗性:	卡那霉素
载体类型:	哺乳细胞表达,CRISPR
高拷贝/低拷贝:	未知
生长条件:	37 ℃
插入基因信息
克隆方法:	TOPO克隆
5' 测序引物	T7
文献引用方法
材料和方法部分:	gRNA_Cloning Vector was a gift from George Church (Addgene plasmid # 41824)
参考文献部分:	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

 

载体质粒图谱和多克隆位点信息

载体简介

 

载体序列

LOCUS       gRNA_Cloning Vector Sequencing Result	3915 bp 	DNA	SYN
DEFINITION  gRNA_Cloning Vector Sequencing Result
ACCESSION   
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