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U6.4-SpCas9 cloning
  • Sequence Map
    Editing disabled on read-only sequence.
    gatggcccactacgtgaaccatcaccctaatcaagttttttggggtcgaggtgccgtaaagcactaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagccggcgaacgtggcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggctaccgggtgatgcacttggtagtgggattagttcaaaaaaccccagctccacggcatttcgtgatttagccttgggatttccctcgggggctaaatctcgaactgcccctttcggccgcttgcaccgctctttccttcccttctttcgctttcctcgcccgcgatcccgcgaccNaeIBsaAIBtgZIDraIIINgoMIVF1 ori20406080100120140160
    caagtgtagcggtcacgctgcgcgtaaccaccacacccgccgcgcttaatgcgccgctacagggcgcgtcccattcgccattcaggctgcgcaactgttgggaagggcgatcggtgcgggcctcttcgctattacgccagctggcgaaagggggatgtgctgcaaggcgattaaggttcacatcgccagtgcgacgcgcattggtggtgtgggcggcgcgaattacgcggcgatgtcccgcgcagggtaagcggtaagtccgacgcgttgacaacccttcccgctagccacgcccggagaagcgataatgcggtcgaccgctttccccctacacgacgttccgctaattcBglIFspIPvuIBsmFILacZ alphaF1 ori180200220240260280300320340
    ttgggtaacgccagggttttcccagtcacgacgttgtaaaacgacggccagtgaattgtaatacgactcactatagggcgaattggagctccaccgcggtggcggccgctctagaactagtggatcccccgggctgcTCTTTGACGAGCGCTATCACGCTGTGCCATTGGTGGTCaacccattgcggtcccaaaagggtcagtgctgcaacattttgctgccggtcacttaacattatgctgagtgatatcccgcttaacctcgaggtggcgccaccgccggcgagatcttgatcacctagggggcccgacgAGAAACTGCTCGCGATAGTGCGACACGGTAACCACCAGAfeIAleIAvaIBmrIEagINotISacISmaISpeIXbaIXmaIBamHIBsoBIBstXISacIITspMIDraIIIEco53kIPU6.4T7Fragme...fwd))M13-fwd360380400420440460480500520
    CTGTTCTGGAAATCCATGGAAATCCCCTCGACGCAGGCCACCCCGACACCACAGCCAAGCGCAGGTCTGCTGGGCTCTGCGAAATCGAATGCAAGTTTAACTGGTGCGCGCGTTCCCTGGACCCTGATTGGCCCATTTTAGAACATTCATGAAGTTGATATACTGCAGATATGCGGACAAGACCTTTAGGTACCTTTAGGGGAGCTGCGTCCGGTGGGGCTGTGGTGTCGGTTCGCGTCCAGACGACCCGAGACGCTTTAGCTTACGTTCAAATTGACCACGCGCGCAAGGGACCTGGGACTAACCGGGTAAAATCTTGTAAGTACTTCAACTATATGACGTCTATACGCBsmINcoIPstIStyIBfuAIBspMIBssHIIPU6.4540560580600620640660680700
    AGAGATAATAAACTAAGCAGTCGGAGGTCGGCCATGCCTCGTGACAGCGGCGTCAACTCCGAGTTCCTGCGGTTCCTGTAGCGCATGAGCATGGGTCGCGTGTGGTGATGACCAGGAACGGCGGCGGCAGGAAGCGGAAGAGGAGGAGGTCAGGAGCAAGAGCtAAGCAGGTAGGTCTCTATTATTTGATTCGTCAGCCTCCAGCCGGTACGGAGCACTGTCGCCGCAGTTGAGGCTCAAGGACGCCAAGGACATCGCGTACTCGTACCCAGCGCACACCACTACTGGTCCTTGCCGCCGCCGTCCTTCGCCTTCTCCTCCTCCAGTCCTCGTTCTCGaTTCGTCCATCCBlpIBfuAIBsaHIBspMIHincIIPU6.4720740760780800820840860
    AGAACGAGGGAGCAGAGTACTTGTGGCTGGGAGGCCACCGGTGCCCTAGCAGGGGCACGCGCCGGTGACGTGAGCAGGAGGAGAAGGCAGTATACGCACTTGACACGGCTTCAGGAGGCGCACAGGAGGCGGAGGCCGCCACGAACCAGCAGAGGAAGCAATAAATTGTCTTACTTCTTGCTCCCTCGTCTCATGAACACCGACCCTCCGGTGGCCACGGGATCGTCCCCGTGCGCGGCCACTGCACTCGTCCTCCTCTTCCGTCATATGCGTGAACTGTGCCGAAGTCCTCCGCGTGTCCTCCGCCTCCGGCGGTGCTTGGTCGTCTCCTTCGTTATTTAACAGAATGAAccIAgeIScaITatIBmgBISgrAISgrAIBstZ17IPU6.48809009209409609801,0001,0201,040
    ATATATATATTTGGCCTATGAGAATTactttgagacgtaccatatgaaccgtctccgttttagagctagaaatagcaagttaaaataaggctagtccgttatcaacttgaaaaagtggcaccgagtcggtgctttttttATCTTTCTAGAAGATCTCCTACAATATTCTCAGCTGTATATATATAAACCGGATACTCTTAAtgaaactctgcatggtatacttggcagaggcaaaatctcgatctttatcgttcaattttattccgatcaggcaatagttgaactttttcaccgtggctcagccacgaaaaaaaTAGAAAGATCTTCTAGAGGATGTTATAAGAGTCGACNdeIXbaIBglIIBsmBIBsmBIEsp3IEsp3ISpCas9 ScaffoldPU6.4Cloning Insert1,0601,0801,1001,1201,1401,1601,1801,2001,220
    CCATGGAAAATCGATGTTCTTCTTTTATTCTCTCAAGATTTTCAGGCTGTATATTAAAACTTATATTAAGAACTATGCTAACCACCTCATCAGGAACCGTTGTAGGTGGCGTGGGTTTTCTTGGCAATCGACTCTCATGAAAACTACGAGCTAAATATTCAATATGTTCCTCTTGGGTACCTTTTAGCTACAAGAAGAAAATAAGAGAGTTCTAAAAGTCCGACATATAATTTTGAATATAATTCTTGATACGATTGGTGGAGTAGTCCTTGGCAACATCCACCGCACCCAAAAGAACCGTTAGCTGAGAGTACTTTTGATGCTCGATTTATAAGTTATACAAGGAGAACClaINcoIStyIBspDI1,2401,2601,2801,3001,3201,3401,3601,3801,400
    ACCAACTTTATTCTGCATTTTTTTTGAACGAggTTTAGAGcaagcttatcgataccgtcgacctcgagggggggcccggtacccagcttttgttccctttagtgagggttaatttcgagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaaTGGTTGAAATAAGACGTAAAAAAAACTTGCTccAAATCTCgttcgaatagctatggcagctggagctcccccccgggccatgggtcgaaaacaagggaaatcactcccaattaaagctcgaaccgcattagtaccagtatcgacaaaggacacactttaacaataggcgagtgttAccIApaIAvaIBcgIClaIKpnISalITliIXhoIBsoBIBspDIPspXIAcc65IHincIIPaeR7IPspOMIHindIIIEcoO109IM13-revT3T3 promoterLacO1,4201,4401,4601,4801,5001,5201,5401,560
    ttccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattggaaggtgtgttgtatgctcggccttcgtatttcacatttcggaccccacggattactcactcgattgagtgtaattaacgcaacgcgagtgacgggcgaaaggtcagccctttggacagcacggtcgacgtaattacttagccggttgcgcgcccctctccgccaaacgcataaccTfiIlac promoter1,5801,6001,6201,6401,6601,6801,7001,7201,740
    gcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgcgcgagaaggcgaaggagcgagtgactgagcgacgcgagccagcaagccgacgccgctcgccatagtcgagtgagtttccgccattatgccaataggtgtcttagtcccctattgcgtcctttcttgtacactcgttttccggtcgttttccggtccttggcatttttccggcgcNspIPciISapITfiIBsaXIBspQIAflIII1,7601,7801,8001,8201,8401,8601,8801,9001,920
    ttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctaacgaccgcaaaaaggtatccgaggcggggggactgctcgtagtgtttttagctgcgagttcagtctccaccgctttgggctgtcctgatatttctatggtccgcaaagggggaccttcgagggagcacgcgagaggacaaggctgggacggcgaatggcctatggacaggcggaBciVIColE1 origin1,9401,9601,9802,0002,0202,0402,0602,0802,100
    ttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcaagagggaagcccttcgcaccgcgaaagagtatcgagtgcgacatccatagagtcaagccacatccagcaagcgaggttcgacccgacacacgtgcttggggggcaagtcgggctggcgacgcggaataggccattgatagcagaactcaggttgggccattctgtgctgaatagApaLIColE1 origin2,1202,1402,1602,1802,2002,2202,2402,260
    gccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggccggtgaccgtcgtcggtgaccattgtcctaatcgtctcgctccatacatccgccacgatgtctcaagaacttcaccaccggattgatgccgatgtgatcttcctgtcataaaccatagacgcgagacgacttcggtcaatggaagcctttttctcaaccatcgagaactaggccgAlwNIColE1 origin2,2802,3002,3202,3402,3602,3802,4002,4202,440
    aaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcaccttttgtttggtggcgaccatcgccaccaaaaaaacaaacgttcgtcgtctaatgcgcgtctttttttcctagagttcttctaggaaactagaaaagatgccccagactgcgagtcaccttgcttttgagtgcaattccctaaaaccagtactctaatagtttttcctagaagtggaColE1 origin2,4602,4802,5002,5202,5402,5602,5802,6002,620
    agatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggtctaggaaaatttaatttttacttcaaaatttagttagatttcatatatactcatttgaaccagactgtcaatggttacgaattagtcactccgtggatagagtcgctagacagataaagcaagtaggtatcaacggactgaggggcagcacatctattgatgctatgccctcccAhdIAmpR2,6402,6602,6802,7002,7202,7402,7602,7802,800
    cttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcggaatggtagaccggggtcacgacgttactatggcgctctgggtgcgagtggccgaggtctaaatagtcgttatttggtcggtcggccttcccggctcgcgtcttcaccaggacgttgaaataggcggaggtaggtcagataattaacaacggcccttcgatctcattcatcaagcBglIBmrIBpmIBsaIBsrDINmeAIIIAmpR2,8202,8402,8602,8802,9002,9202,9402,960
    ccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaaggtcaattatcaaacgcgttgcaacaacggtaacgatgtccgtagcaccacagtgcgagcagcaaaccataccgaagtaagtcgaggccaagggttgctagttccgctcaatgtactagggggtacaacacgttttttcgccaatcgaggaagccaggaggctagcaacagtcttAclIFspIPvuIBsrDIAmpR2,9803,0003,0203,0403,0603,0803,1003,1203,140
    gtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccattcaaccggcgtcacaatagtgagtaccaataccgtcgtgacgtattaagagaatgacagtacggtaggcattctacgaaaagacactgaccactcatgagttggttcagtaagactcttatcacatacgccgctggctcaacgagaacgggccgcagttatgccctattatgBcgIScaITatIBsaHIAmpR3,1603,1803,2003,2203,2403,2603,2803,3003,320
    cgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggagcgcggtgtatcgtcttgaaattttcacgagtagtaaccttttgcaagaagccccgcttttgagagttcctagaatggcgacaactctaggtcaagctacattgggtgagcacgtgggttgactagaagtcgtagaaaatgaaagtggtcgcaaagacccactcgtttttgtcctAclIXmnIApaLIAmpR3,3403,3603,3803,4003,4203,4403,4603,4803,500
    aggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaatccgttttacggcgttttttcccttattcccgctgtgcctttacaacttatgagtatgagaaggaaaaagttataataacttcgtaaatagtcccaataacagagtactcgcctatgtataaacttacataaatctttttatttgtttatccccaaggcgcgtgtaaaggggcttBciVI3,5203,5403,5603,5803,6003,6203,6403,660
    aagtgccacctaaattgtaagcgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcattttttaaccaataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttgagtgttgttccagtttggaacaagagtccactattattcacggtggatttaacattcgcaattataaaacaattttaagcgcaatttaaaaacaatttagtcgagtaaaaaattggttatccggctttagccgttttagggaatatttagttttcttatctggctctatcccaactcacaacaaggtcaaaccttgttctcaggtgataatAloIApoIApoIPsiIBsaXICspCIF1 ori3,6803,7003,7203,7403,7603,7803,8003,8203,840
    aagaacgtggactccaacgtcaaagggcgaaaaaccgtctatcagggcttcttgcacctgaggttgcagtttcccgctttttggcagatagtcccgF1 ori3,8603,880
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    Editing disabled on read-only sequence.
    3898 bp27503000325035003750250500750AmpRPU6.4pBluKSP_SaCas9\Scaffold\cloning REV (DNA)p046_PU6.4_(pJET,_842) FWD (DNA)

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    3898