LOCUS       pT3TS-hypa-nCas9        7486 bp    DNA     circular      8-APR-2019
DEFINITION  pT3TS-nCas9n.
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTDATE|-11918371|
COMMENT     VNTDBDATE|-12761982|
COMMENT     LSOWNER|
COMMENT     VNTNAME|pT3TS-hypa-nCas9n-pA|
COMMENT     VNTAUTHORNAME|Demo User|
COMMENT     VNTOAUTHORNAME|UNKNOWN|
FEATURES             Location/Qualifiers
     source          1..7486
                     /organism="pT3TS-nCas9n"
                     /mol_type="other DNA"
                     /vntifkey="98"
     promoter        1..20
                     /vntifkey="30"
                     /label=T3_promoter
     CDS             108..4286
                     /vntifkey="4"
                     /label=nCas9n
     misc_feature    120..140
                     /vntifkey="21"
                     /label=SV40\NLS
     misc_feature    120..143
                     /vntifkey="21"
                     /label=SV40\NLS
     misc_feature    126..140
                     /vntifkey="21"
                     /label=NLS
     promoter        complement(4676..4694)
                     /vntifkey="30"
                     /label=T7_promoter
     misc_feature    4698..4853
                     /vntifkey="21"
                     /label=lacZ_a
     promoter        complement(4701..4717)
                     /vntifkey="30"
                     /label=M13_forward20_primer
     misc_feature    complement(4710..4732)
                     /vntifkey="21"
                     /label=M13_pUC_fwd_primer
     rep_origin      complement(4990..5296)
                     /vntifkey="33"
                     /label=f1_origin
     promoter        5375..5403
                     /vntifkey="30"
                     /label=AmpR_promoter
     gene            5445..6305
                     /gene="Ampicillin"
                     /vntifkey="60"
     CDS             5445..6305
                     /vntifkey="4"
                     /label=ORF\frame\3
     rep_origin      6460..7079
                     /vntifkey="33"
                     /label=pBR322_origin
     promoter        7388..7417
                     /vntifkey="30"
                     /label=lac_promoter
     misc_feature    7431..7453
                     /vntifkey="21"
                     /label=M13_pUC_rev_primer
     promoter        7452..7470
                     /vntifkey="30"
                     /label=M13_reverse_primer
     misc_feature    4293..4442
                     /vntifkey="21"
                     /label=A69-polyA
     misc_feature    2220..2240
                     /vntifkey="21"
                     /label=hypaCas9_mutated_region
     misc_feature    2220..2222
                     /vntifkey="21"
                     /label=N692A
     misc_feature    2226..2228
                     /vntifkey="21"
                     /label=M694A
     misc_feature    2229..2231
                     /vntifkey="21"
                     /label=Q695A
     misc_feature    2238..2240
                     /vntifkey="21"
                     /label=H698A
     misc_feature    147..4283
                     /vntifkey="21"
                     /label=Cas9_CDS
     misc_feature    3546..3548
                     /vntifkey="21"
                     /label=D1134
     misc_feature    3795..3797
                     /vntifkey="21"
                     /label=G1217
     misc_feature    4146..4148
                     /vntifkey="21"
                     /label=R1334
     misc_feature    4152..4154
                     /vntifkey="21"
                     /label=T1336
BASE COUNT     2264 a      1716 c      1867 g      1639 t 
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga 
       61 agctcagaat aaacgctcaa ctttggcaga tctaactcga cgccaccatg gcttctccac 
      121 ctaagaagaa gagaaaggtg ggaagcatgg ataagaagta tagcatcggc ctggatattg 
      181 gaactaactc cgtgggttgg gcagtgatta cagacgacta caaggtccct agcaagaaat 
      241 ttaaggtgct gggtaacacc gacaggcaca gcatcaagaa aaatctgatt ggagccctgc 
      301 tgttcggttc tggagagact gccgaagcaa cacgcctgaa aagaacagca agaaggcgct 
      361 ataccagaag gaagaataga atctgttacc tgcaggagat tttctctaac gaaatggcta 
      421 aggtggacga ttcattcttt cataggctgg aggaaagttt cctggtcgag gaagataaga 
      481 aacacgagcg ccatcctatc tttggaaaca ttgtggacga ggtcgcctat cacgaaaaat 
      541 acccaaccat ctatcatctg cgcaagaaac tggctgactc tactgataaa gccgacctga 
      601 gactgatcta tctggctctg gcccacatga ttaagttcag gggtcatttt ctgatcgagg 
      661 gcgatctgaa ccccgacaat tccgatgtgg acaagctgtt catccagctg gtccagattt 
      721 acaatcagct gtttgaggaa aaccctatta atgcttccag agtggacgca aaagctatcc 
      781 tgtcagccag gctgtccaag tcacgcagac tggagaacct gattgcacag ctgcccggag 
      841 aaaagaggaa cggtctgttt ggaaatctga tcgctctgag tctgggcctg actcctaact 
      901 tcaaaagcaa ttttgatctg gctgaggacg ccaaactgca gctgtcaaag gacacatatg 
      961 acgatgacct ggataacctg ctggcacaga tcggagatca gtacgctgac ctgttcctgg 
     1021 ctgccaaaaa tctgtccgac gcaatcctgc tgtcagatat tctgagagtg aacagcgaga 
     1081 ttacaaaagc tcctctgagt gccagcatga tcaagagata tgacgagcac catcaggatc 
     1141 tgaccctgct gaaggctctg gtcaggcagc agctgccaga gaagtacaag gaaattttct 
     1201 ttgatcagtc caagaacggc tacgccggtt atatcgacgg aggcgcatca caggaggaat 
     1261 tctacaagtt tatcaaacct attctggaga agatggacgg aactgaggaa ctgctggtga 
     1321 aactgaatag agaggacctg ctgaggaagc agcgcacatt tgataacggt tccatcccac 
     1381 accagattca tctgggagag ctgcacgcta tcctgaggcg ccaggaagac ttctacccct 
     1441 ttctgaaaga taaccgcgag aagatcgaaa aaattctgac cttcagaatc ccttactatg 
     1501 tgggtccact ggctcgcgga aacagcagat ttgcctggat gactcgcaaa tccgaggaaa 
     1561 ccattactcc ttggaacttc gaggaagtgg tcgataaggg cgcctctgca cagtccttca 
     1621 tcgagagaat gactaatttt gacaaaaacc tgcccaatga gaaagtgctg cctaagcact 
     1681 ccctgctgta cgagtatttc actgtctata acgaactgac aaaggtgaaa tacgtcaccg 
     1741 agggcatgag aaagccagcc ttcctgtcag gagagcagaa gaaagcaatc gtggatctgc 
     1801 tgtttaaaac caacaggaaa gtgactgtca agcagctgaa ggaggactac ttcaagaaaa 
     1861 ttgaatgctt cgattccgtg gagatcagcg gagtcgaaga cagatttaac gcaagcctgg 
     1921 gcgcttacca cgatctgctg aagatcatta aggataaaga cttcctggac aacgaggaaa 
     1981 atgaggatat cctggaagac attgtgctga cactgaccct gtttgaggac agaggaatga 
     2041 tcgaggaaag actgaaaacc tatgctcatc tgttcgatga caaggtgatg aaacagctga 
     2101 agagaaggcg ctacactggc tggggtagac tgagcaggaa gctgatcaac ggcattaggg 
     2161 ataaacagtc aggaaagaca atcctggact ttctgaaaag tgatggcttc gccaaccgcg 
     2221 cttttgcggc actgattgcc gatgacagtc tgaccttcaa agaggacatc cagaaggctc 
     2281 aggtgtctgg acagggccac tccctgcatg agcagattgc aaacctggct ggaagcccag 
     2341 ccatcaagaa aggcattctg cagacagtga aaatcgtcga tgagctggtg aaagtcatgg 
     2401 gccataagcc cgaaaacatc gtgattgaga tggctcgcga aaatcagaca acccagaagg 
     2461 gtcagaagaa cagtagagag aggatgaaaa gaatcgagga aggcattaag gagctgggta 
     2521 gccagatcct gaaagagcac ccagtggaaa acacacagct gcagaatgag aagctgtatc 
     2581 tgtactatct gcagaatgga agagatatgt acgtggacca ggagctggat attaacaggc 
     2641 tgtctgatta cgacgtggat catatcgtcc cccagagttt catcaaagat gacagcattg 
     2701 acaacaaggt gctgaccagg tccgacaaaa acagaggaaa atcagataat gtccctagtg 
     2761 aggaagtggt caagaaaatg aagaactact ggagacagct gctgaatgcc aaactgatca 
     2821 ctcagaggaa gtttgataac ctgacaaaag cagagcgcgg tggactgtca gaactggaca 
     2881 aagctggatt catcaagagg cagctggtgg aaacacgcca gatcactaaa cacgtcgcac 
     2941 agattctgga tagtcgcatg aacacaaagt acgatgagaa tgacaaactg atcagagaag 
     3001 tgaaggtcat taccctgaag agtaaactgg tcagcgactt taggaaagat ttccagtttt 
     3061 ataaggtccg cgagattaac aattatcacc atgcccatga cgcatacctg aacgccgtgg 
     3121 tcggtaccgc actgatcaag aaatacccaa aactggagag cgaattcgtg tacggagact 
     3181 ataaggtgta cgatgtcaga aaaatgatcg ccaagtccga gcaggaaatt ggaaaagcta 
     3241 ctgccaagta tttcttttac tcaaacatca tgaatttctt taagacagag atcaccctgg 
     3301 ccaatggaga aatccgcaaa aggcccctga ttgagacaaa cggagagaca ggcgaaatcg 
     3361 tgtgggacaa aggcagagat tttgcaaccg tgaggaaggt cctgagcatg cctcaagtga 
     3421 atatcgtcaa gaaaactgag gtgcagacag gcggtttctc aaaagaaagt attctgccaa 
     3481 aacgcaactc tgataagctg atcgctagaa agaaagactg ggaccctaag aagtatggag 
     3541 gctttgactc tcccactgtg gcatactccg tcctggtggt cgctaaggtg gagaagggca 
     3601 aaagcaagaa actgaaatct gtcaaggagc tgctgggtat cacaattatg gagagaagct 
     3661 ctttcgagaa gaacccaatc gattttctgg aggccaaagg ttataaggaa gtgaagaaag 
     3721 acctgatcat taaactgccc aagtacagtc tgtttgagct ggaaaacggc aggaaacgca 
     3781 tgctggcaag cgctggagag ctgcagaaag gcaatgaact ggccctgcct tctaagtacg 
     3841 tgaacttcct gtatctggca agccactacg agaagctgaa aggatctcca gaggataacg 
     3901 aacagaaaca gctgtttgtg gagcagcaca agcattatct ggacgagatc attgaacaga 
     3961 ttagcgagtt ctctaaaaga gtgatcctgg ccgacgcaaa tctggataag gtcctgtctg 
     4021 cttacaacaa acacagagat aagcccatca gggagcaggc cgaaaatatc attcatctgt 
     4081 tcactctgac aaacctgggc gcacctgcag ctttcaagta cttcgacact acaatcgata 
     4141 gaaagaggta cacctccact aaggaggtgc tggacgctac actgatccat cagagtatta 
     4201 ccggcctgta cgaaacaagg attgacctgt ctcagctggg tggcgactca ccggtgagat 
     4261 ctcctaagaa gaagagaaag gtgtgaccgc gggcaccaaa gaacctgtaa acgttatctt 
     4321 tttaaattga atgtgcacaa ataaaagttt ggaaaagaaa aaaaaaaaaa aaaaaaaaaa 
     4381 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa actcgaggta gcatgctcta 
     4441 gaccgcggat ctggttacca ctaaaccagc ctcaagaaca cccgaatgga gtctctaagc 
     4501 tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc cattcgtatc 
     4561 tgctcctaat aaaaagaaag tttcttcaca ttctaaaaaa aaaaaaaaaa aaaaaaaaaa 
     4621 aaaccccccc ccccccctgc agggccaagt cggcctctag aggtacccaa ttcgccctat 
     4681 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 
     4741 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 
     4801 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa 
     4861 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 
     4921 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 
     4981 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 
     5041 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 
     5101 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 
     5161 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 
     5221 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 
     5281 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc 
     5341 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 
     5401 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 
     5461 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 
     5521 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 
     5581 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 
     5641 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 
     5701 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 
     5761 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 
     5821 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 
     5881 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 
     5941 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 
     6001 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 
     6061 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 
     6121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 
     6181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 
     6241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 
     6301 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 
     6361 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 
     6421 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 
     6481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 
     6541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 
     6601 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 
     6661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 
     6721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 
     6781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 
     6841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 
     6901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 
     6961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 
     7021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 
     7081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 
     7141 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 
     7201 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 
     7261 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 
     7321 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac 
     7381 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac 
     7441 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcga 
//
