LOCUS       pX458-DsRed2        9253 bp    DNA     circular     23-MAY-2018
SOURCE      
  ORGANISM  
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
COMMENT     VNTDATE|-12301197|
COMMENT     VNTDBDATE|-12301198|
COMMENT     LSOWNER|
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COMMENT     VNTAUTHORNAME|ZHANG Lichen|
COMMENT     VNTOAUTHORNAME|ZHANG LICHEN|
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FEATURES             Location/Qualifiers
     promoter        10..242
                     /vntifkey="29"
                     /label=hU6
     CDS             7573..8433
                     /vntifkey="4"
                     /label=Amp
     enhancer        444..713
                     /vntifkey="9"
                     /label=CAG_enhancer
     misc_feature    1254..1319
                     /vntifkey="21"
                     /label=FLAG
     misc_feature    268..343
                     /vntifkey="21"
                     /label=gRNA\scaffold
     terminator      344..349
                     /vntifkey="43"
                     /label=polIII\terminator
     CDS             1371..5471
                     /vntifkey="4"
                     /label=Cas9
     misc_signal     1320..1370
                     /vntifkey="87"
                     /label=NLS
     misc_signal     5472..5519
                     /vntifkey="87"
                     /label=NLS
     misc_feature    5535..5588
                     /vntifkey="21"
                     /label=T2A
     polyA_signal    6300..6507
                     /vntifkey="25"
                     /label=bGH\poly(A)\signal
     rep_origin      6731..7186
                     /vntifkey="33"
                     /label=f1\ori
     promoter        7468..7572
                     /vntifkey="30"
                     /label=AmpR\promoter
     CDS             5589..6266
                     /vntifkey="4"
                     /label=DsRed2
     mutation        6005..6005
                     /vntifkey="62"
                     /label=g417a
                     /note="417th of DeRed2 g mutate to a"
     primer          5328..5347
                     /vntifkey="27"
                     /label=Fwd\sequencing
     primer          complement(6369..6388)
                     /vntifkey="27"
                     /label=Rev\sequencing
     primer          328..349
                     /vntifkey="27"
                     /label=IVT-R
BASE COUNT     2387 a      2480 c      2534 g      1852 t 
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct 
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac 
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt 
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 
      721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 
      781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 
      841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 
      901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 
      961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc 
     1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg 
     1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag 
     1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac 
     1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata 
     1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga 
     1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt 
     1381 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt 
     1441 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga 
     1501 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga 
     1561 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga 
     1621 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct 
     1681 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg 
     1741 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca 
     1801 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc 
     1861 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt 
     1921 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg 
     1981 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc 
     2041 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga 
     2101 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc 
     2161 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc 
     2221 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca 
     2281 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat 
     2341 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg 
     2401 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg 
     2461 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg 
     2521 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct 
     2581 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc 
     2641 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga 
     2701 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga 
     2761 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg 
     2821 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg 
     2881 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga 
     2941 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga 
     3001 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga 
     3061 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag 
     3121 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg 
     3181 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac 
     3241 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg 
     3301 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga 
     3361 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt 
     3421 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta 
     3481 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg 
     3541 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg 
     3601 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca 
     3661 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg 
     3721 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc 
     3781 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg 
     3841 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga 
     3901 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg 
     3961 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc 
     4021 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga 
     4081 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc 
     4141 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg 
     4201 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg 
     4261 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc 
     4321 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg 
     4381 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga 
     4441 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact 
     4501 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga 
     4561 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga 
     4621 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct 
     4681 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg 
     4741 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg 
     4801 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg 
     4861 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca 
     4921 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg 
     4981 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg 
     5041 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc 
     5101 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact 
     5161 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg 
     5221 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc 
     5281 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca 
     5341 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg 
     5401 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc 
     5461 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg 
     5521 aattcggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc 
     5581 ctggcccaat ggcctcctcc gagaacgtca tcaccgagtt catgcgcttc aaggtgcgca 
     5641 tggagggcac cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 
     5701 acgagggcca caacaccgtg aagctgaagg tgaccaaggg cggccccctg cccttcgcct 
     5761 gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg 
     5821 acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 
     5881 acttcgagga cggcggcgtg gcgaccgtga cccaggactc ctccctgcag gacggctgct 
     5941 tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc gtgatgcaga 
     6001 agaaaaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga 
     6061 agggcgagac ccacaaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca 
     6121 agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac gtggacgcca 
     6181 agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac gagcgcaccg 
     6241 agggccgcca ccacctgttc ctgtaggaat tctaactaga gctcgctgat cagcctcgac 
     6301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 
     6361 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 
     6421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 
     6481 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc 
     6541 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg 
     6601 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg 
     6661 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag 
     6721 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 
     6781 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 
     6841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 
     6901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca 
     6961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 
     7021 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct 
     7081 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 
     7141 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc 
     7201 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct 
     7261 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc 
     7321 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag 
     7381 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg 
     7441 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 
     7501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 
     7561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca 
     7621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat 
     7681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag 
     7741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc 
     7801 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct 
     7861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca 
     7921 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt 
     7981 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat 
     8041 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt 
     8101 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta 
     8161 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 
     8221 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 
     8281 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 
     8341 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 
     8401 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 
     8461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 
     8521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 
     8581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 
     8641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 
     8701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 
     8761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 
     8821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 
     8881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 
     8941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 
     9001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 
     9061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 
     9121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 
     9181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 
     9241 tttgctcaca tgt 
//
