pCas-Guide-Nickase

pCas-Guide-Nickase

pCas-Guide-Nickase载体基本信息：

pCas-Guide-Nickase载体质粒图谱和多克隆位点信息：

pCas-Guide-Nickase载体简介：

Expresses human codon-optimized Cas9 nickase D10A mutant, C-terminal Myc-DDK tagged

BamH I and BsmB I can be used for genomic target sequence cloning

Expresses gRNA containing the inserted target sequence and the gRNA scaffold

CF3 primer can be used to sequence the target sequence after cloning

pCas-Guide-Nickase载体序列：

ORIGIN

        1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc

       61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt

      121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa

      181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt

      241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt

      301 tcttgggttt atatatcttg tggaaaggac gcgggatcca ctggaccagg cagcagcgtc

      361 agaagacttt tttggaacgt ctcgttttag agctagaaat agcaagttaa aataaggcta

      421 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttggtg tacatttata

      481 ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt

      541 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta

      601 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga

      661 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt

      721 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta

      781 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg

      841 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt

      901 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc

      961 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat

     1021 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct

     1081 atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc actatagggc

     1141 ggccgggaat tcgtcgactg gaaccggtac cgaggagatc tgccgccgcg atcgccatgg

     1201 ataagaaata ctcaatagga ctggctattg gcacaaatag cgtcggatgg gctgtgatca

     1261 ctgatgaata taaggttcct tctaaaaagt tcaaggttct gggaaataca gaccgccaca

     1321 gtatcaaaaa aaatcttata ggggctcttc tgtttgacag tggagagaca gccgaagcta

     1381 ctagactcaa acggacagct aggagaaggt atacaagacg gaagaatagg atttgttatc

     1441 tccaggagat tttttcaaat gagatggcca aagtggatga tagtttcttt catagacttg

     1501 aagagtcttt tttggtggaa gaagacaaga agcatgaaag acatcctatt tttggaaata

     1561 tagtggatga agttgcttat cacgagaaat atccaactat ctatcatctg agaaaaaaat

     1621 tggtggattc tactgataaa gccgatttgc gcctgatcta tttggccctg gcccacatga

     1681 ttaagtttag aggtcatttt ttgattgagg gcgatctgaa tcctgataat agtgatgtgg

     1741 acaaactgtt tatccagttg gtgcaaacct acaatcaact gtttgaagaa aaccctatta

     1801 acgcaagtgg agtggatgct aaagccattc tttctgcaag attgagtaaa tcaagaagac

     1861 tggaaaatct cattgctcag ctccccggtg agaagaaaaa tggcctgttt gggaatctca

     1921 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg

     1981 ctaaactcca gctttcaaaa gatacttacg atgatgatct ggataatctg ttggctcaaa

     2041 ttggagatca atatgctgat ttgtttttgg cagctaagaa tctgtcagat gctattctgc

     2101 tttcagacat cctgagagtg aatactgaaa taactaaggc tcccctgtca gcttcaatga

     2161 ttaaacgcta cgatgaacat catcaagact tgactcttct gaaagccctg gttagacaac

     2221 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt

     2281 atattgatgg cggcgcaagc caagaagaat tttataaatt tatcaaacca attctggaaa

     2341 aaatggatgg tactgaggaa ctgttggtga aactgaatag agaagatttg ctgcgcaagc

     2401 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta

     2461 ttttgagaag acaagaagac ttttatccat ttctgaaaga caatagagag aagattgaaa

     2521 aaatcttgac ttttaggatt ccttattatg ttggtccatt ggccagaggc aatagtaggt

     2581 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg

     2641 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc

     2701 ttccaaatga aaaagtgctg ccaaaacata gtttgcttta tgagtatttt accgtttata

     2761 acgaattgac aaaggtcaaa tatgttactg aaggaatgag aaaaccagca tttctttcag

     2821 gtgaacagaa gaaagccatt gttgatctgc tcttcaaaac aaataggaaa gtgaccgtta

     2881 agcaactgaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag

     2941 gagttgaaga tagatttaat gcttcactgg gtacatacca tgatttgctg aaaattatta

     3001 aagataaaga ttttttggat aatgaagaaa atgaagacat cctggaggat attgttctga

     3061 cattgaccct gtttgaagat agggagatga ttgaggaaag acttaaaaca tacgctcacc

     3121 tctttgatga taaggtgatg aaacagctta aaagacgcag atatactggt tggggaaggt

     3181 tgtccagaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atactggatt

     3241 ttttgaaatc agatggtttt gccaatcgca attttatgca gctcatccat gatgatagtt

     3301 tgacatttaa agaagacatc caaaaagcac aagtgtctgg acaaggcgat agtctgcatg

     3361 aacatattgc aaatctggct ggtagccctg ctattaaaaa aggtattctc cagactgtga

     3421 aagttgttga tgaattggtc aaagtgatgg ggcggcataa gccagaaaat atcgttattg

     3481 aaatggcaag agaaaatcag acaactcaaa agggccagaa aaattccaga gagaggatga

     3541 aaagaatcga agaaggtatc aaagaactgg gaagtcagat tcttaaagag catcctgttg

     3601 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagata

     3661 tgtatgtgga ccaagaactg gatattaata ggctgagtga ttatgatgtc gatcacattg

     3721 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcctgacc aggtctgata

     3781 aaaatagagg taaatccgat aacgttccaa gtgaagaagt ggtcaaaaag atgaaaaact

     3841 attggagaca acttctgaac gccaagctga tcactcaaag gaagtttgat aatctgacca

     3901 aagctgaaag aggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg

     3961 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta

     4021 aatacgatga aaatgataaa cttattagag aggttaaagt gattaccctg aaatctaaac

     4081 tggtttctga cttcagaaaa gatttccaat tctataaagt gagagagatt aacaattacc

     4141 atcatgccca tgatgcctat ctgaatgccg tcgttggaac tgctttgatt aagaaatatc

     4201 caaaacttga aagcgagttt gtctatggtg attataaagt ttatgatgtt aggaaaatga

     4261 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa gtatttcttt tactctaata

     4321 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc

     4381 tgatcgaaac taatggggaa actggagaaa ttgtctggga taaagggaga gattttgcca

     4441 cagtgcgcaa agtgttgtcc atgccccaag tcaatatcgt caagaaaaca gaagtgcaga

     4501 caggcggatt ctctaaggag tcaattctgc caaaaagaaa ttccgacaag ctgattgcta

     4561 ggaaaaaaga ctgggaccca aaaaaatatg gtggttttga tagtccaacc gtggcttatt

     4621 cagtcctggt ggttgctaag gtggaaaaag ggaaatccaa gaagctgaaa tccgttaaag

     4681 agctgctggg gatcacaatt atggaaagaa gttcctttga aaaaaatccc attgactttc

     4741 tggaagctaa aggatataag gaagttaaaa aagacctgat cattaaactg cctaaatata

     4801 gtctttttga gctggaaaac ggtaggaaac ggatgctggc tagtgccgga gaactgcaaa

     4861 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tctgtatctg gctagtcatt

     4921 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc

     4981 ataagcatta tctggatgag attattgagc aaatcagtga attttctaag agagttattc

     5041 tggcagatgc caatctggat aaagttctta gtgcatataa caaacataga gacaaaccaa

     5101 taagagaaca agcagaaaat atcattcatc tgtttacctt gaccaatctt ggagcacccg

     5161 ctgcttttaa atactttgat acaacaattg ataggaaaag atatacctct acaaaagaag

     5221 ttctggatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt

     5281 tgagtcagct gggaggtgac cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc

     5341 ggaaagtgga cacgcgtacg cggccgctcg agcagaaact catctcagaa gaggatctgg

     5401 cagcaaatga tatcctggat tacaaggatg acgacgataa ggtttaaacg gccggccgcg

     5461 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct

     5521 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt

     5581 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta

     5641 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca

     5701 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg

     5761 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc

     5821 taatttttgt ttttttggta gaggcggggt ttcaccatat tggccaggct ggtctccaac

     5881 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga

     5941 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc

     6001 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt

     6061 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg

     6121 ccagcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg

     6181 ggataacgca ggaaagaaca tgtccgtaaa aaggccgcgt tgctggcgtt tttccatagg

     6241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg

     6301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt

     6361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt

     6421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc

     6481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt

     6541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt

     6601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc

     6661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa

     6721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt

     6781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct

     6841 acggggtctg acgctcagtg gaacgacgcg taactcacgt taagggattt tggtcatgag

     6901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat

     6961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc

     7021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat

     7081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc

     7141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag

     7201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag

     7261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt

     7321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg

     7381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt

     7441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc

     7501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc

     7561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa

     7621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg

     7681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc

     7741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag

     7801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt

     7861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgatga tattatttta

     7921 tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcca ggaaacagct

     7981 atgaccatgt aatacgactc actatagggg atatcagctg gatggcagtt aac

//

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