LOCUS       (selected)_pAAV-U6-Pet1 5294 bp ds-DNA     circular     17-FEB-2022
DEFINITION  .
COMMENT     Primer design for gRNA Vipr1 3종: 1.1_INF_Mlu1-6-U6_cnF
            ctgcggccgcacgcgtCGTGTAgagggcctatttcccatg 1.1_MG_VIPR1-1-U6_cnR :
            TCGGACCCGCGGTGAGTGTGC GGTGTTTCGTCCTTT 1.2_MG_VIPR1-1-Sca_cnF :
            GCACACTCACCGCGGGTCCGA gttttagagctaGAA 1.2_INF_5-Asc1-5-Sca_cnR
            gcagtctagaACCGGccaggggcgcgccgtcgcTAAAAAAgcaccgactcgg
            2.1_INF_Mlu1-5-Asc1-5-U6_cnF
            ctgcggccgcacgcgtgcgacggcgcgcccctgggagggcctatttcccatg
            2.1_MG_VIPR1-2-U6_cnR : CTTGGGGCCAGGTGGTAGTC GGTGTTTCGTCCTTT
            2.2_MG_VIPR1-2-Sca_cnF : GACTACCACCTGGCCCCAAG gttttagagctaGAA
            2.2_INF-4-Nhe1-5-Sca_cnR
            gcagtctagaACCGGgcatgctagccaccaTAAAAAAgcaccgactcgg
            3.1_INF-Mlu1-5-Nhe1-4-U6_cnF
            ctgcggccgcacgcgttggtggctagcatgcgagggcctatttcccatg
            3.1_MG_VIPR1-3-U6_cnR : CCGCCGTCTTCATCAAGGATC GGTGTTTCGTCCTTT
            3.2_MG_VIPR1-3-Sca_cnF : GATCCTTGATGAAGACGGCGG gttttagagctaGAA
            3.2_INF_Age1_cnR gcagtctagaACCGGT
FEATURES             Location/Qualifiers
     misc_feature    1..141
                     /label="L-ITR"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     misc_feature    151..155
                     /label="MluI"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     primer          156..177
                     /label="Seq_sgRNA"
                     /note="sequence: gagggcctatttcccatgattc"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     promoter        156..404
                     /label="Human U6 Promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    224..992
                     /label="hU6 sequencing"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    402..404
                     /label="5'"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     misc_feature    405..428
                     /label="Pet1-gRNA#1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    429..431
                     /label="3'"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     misc_feature    429..510
                     /label="chimeric RNA backbone 1 (SapI)"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     misc_feature    511..516
                     /label="XbaI"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     misc_feature    511..992
                     /label="hSyn"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     primer          complement(579..598)
                     /label="hSyn_seqR"
                     /note="sequence: gtcggtcgtcaggtaggcac"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     exon            1020..1724
                     /label="mCherry"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1020..1724
                     /label="hSyn-F (2 mis)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             1749..1997
                     /label="KASH domain of Nesprin2"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     misc_feature    2006..2019
                     /label="WPRE"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     polyA_signal    2039..2517
                     /label="hGH polyA"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_fwdcolor="#ff0080"
     misc_feature    2557..2697
                     /label="R-ITR"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     rep_origin      2789..3095
                     /label="f1 Origin"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3614..4471
                     /label="AmpR"
                     /ApEinfo_revcolor="#0080ff"
                     /ApEinfo_fwdcolor="#0080ff"
     rep_origin      4622..5289
                     /label="pUC ori"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgaggg cctatttccc atgattcctt
      181 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
      241 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
      301 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
      361 tcgatttctt ggctttatat atcttGTGGA AAGGACGAAA CACCGCCGCG CCACCTCGTC
      421 CGGGTCGGgt tttagagcta GAAAtagcaa gttaaaataa ggctagtccg ttatcaactt
      481 gaaaaagtgg caccgagtcg gtgcTTTTTT tctagactgc agagggccct gcgtatgagt
      541 gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc
      601 cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag
      661 ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac
      721 agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct
      781 gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc
      841 cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga
      901 ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg
      961 agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg atcctctaga gTCGACatgg
     1021 tgagcaaggg cgaggaggat aacatggcca tcatcGagga gttcatgcgc ttcaaggtgc
     1081 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1141 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     1201 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     1261 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     1321 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     1381 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1441 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1501 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1561 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1621 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     1681 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagGAATTC tccggactca
     1741 gatctcgaga ggaggaggag gagacagaca gcaggatgcc ccacctcgac agccccggca
     1801 gctcccagcc gagacgctcc ttcctctcaa gggtgatcag ggcagcgcta ccgttgcagc
     1861 tgcttctgct gctgctgctg ctcctggcct gcctgctacc tgcctctgaa gatgactaca
     1921 gctgcaccca ggccaacaac tttgcccgat ccttctaccc catgctgcgg tacaccaacg
     1981 ggccacctcc cacctagGAT atcaagctta tcgataccga gcgctgctcg agagatctac
     2041 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     2101 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     2161 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     2221 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     2281 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     2341 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     2401 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     2461 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     2521 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2581 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2641 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2701 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2761 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3181 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3241 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3301 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3361 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5281 ttttgctcac atgt
//