pSpCas9(BB)-2A-Puro (PX459) V2.0

LOCUS       pSpCas9(BB)-2A-Puro (PX459) V2.0	9175 bp 	DNA	SYN
DEFINITION  pSpCas9(BB)-2A-Puro (PX459) V2.0
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..9175
                     /organism="pSpCas9(BB)-2A-Puro (PX459) V2.0"
                     /mol_type="other DNA"
     promoter        10..242
                     /label="hU6_promoter"
     misc_feature    172..191
                     /label="LKO1_5_primer"
     misc_feature    444..713
                     /label="CAG_enhancer"
     CDS             complement(1230..3887)
                     /label="ORF frame 3"
     CDS             1251..6188
                     /label="ORF frame 3"
     misc_feature    1254..1274
                     /label="FLAG"
     misc_feature    1275..1295
                     /label="FLAG"
     misc_feature    1296..1319
                     /label="FLAG"
     misc_feature    1326..1346
                     /label="SV40 NLS"
     misc_feature    1326..1349
                     /label="SV40 NLS"
     misc_feature    1332..1346
                     /label="NLS"
     gene            5589..6188
                     /label="puro (variant)"
                     /gene="puro (variant)"
     misc_feature    complement(6216..6233)
                     /label="BGH_rev_primer"
     terminator      6219..6422
                     /label="bGH_PA_terminator"
     rep_origin      6670..6976
                     /label="f1_origin"
     misc_feature    7244..7266
                     /label="pGEX_3_primer"
     promoter        7425..7453
                     /label="AmpR_promoter"
     gene            7495..8355
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             7495..8355
                     /label="ORF frame 1"
     rep_origin      8510..9129
                     /label="pBR322_origin"
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 GACCGAGTAC AAGCCCACGG TGCGCCTCGC CACCCGCGAC GACGTCCCCA
 5641 GGGCCGTACG CACCCTCGCC GCCGCGTTCG CCGACTACCC CGCCACGCGC CACACCGTCG
 5701 ATCCGGACCG CCACATCGAG CGGGTCACCG AGCTGCAAGA ACTCTTCCTC ACGCGCGTCG
 5761 GGCTCGACAT CGGCAAGGTG TGGGTCGCGG ACGACGGCGC CGCGGTGGCG GTCTGGACCA
 5821 CGCCGGAGAG CGTCGAAGCG GGGGCGGTGT TCGCCGAGAT CGGCCCGCGC ATGGCCGAGT
 5881 TGAGCGGTTC CCGGCTGGCC GCGCAGCAAC AGATGGAAGG CCTCCTGGCG CCGCACCGGC
 5941 CCAAGGAGCC CGCGTGGTTC CTGGCCACCG TCGGAGTCTC GCCCGACCAC CAGGGCAAGG
 6001 GTCTGGGCAG CGCCGTCGTG CTCCCCGGAG TGGAGGCGGC CGAGCGCGCC GGGGTGCCCG
 6061 CCTTCCTGGA GACCTCCGCG CCCCGCAACC TCCCCTTCTA CGAGCGGCTC GGCTTCACCG
 6121 TCACCGCCGA CGTCGAGGTG CCCGAAGGAC CGCGCACCTG GTGCATGACC CGCAAGCCCG
 6181 GTGCCTGAga attctaaCTA GAGCTCGCTG ATCAGCCTCG ACTGTGCCTT CTAGTTGCCA
 6241 GCCATCTGTT GTTTGCCCCT CCCCCGTGCC TTCCTTGACC CTGGAAGGTG CCACTCCCAC
 6301 TGTCCTTTCC TAATAAAATG AGGAAATTGC ATCGCATTGT CTGAGTAGGT GTCATTCTAT
 6361 TCTGGGGGGT GGGGTGGGGC AGGACAGCAA GGGGGAGGAT TGGGAAGAgA ATAGCAGGCA
 6421 TGCTGGGGAg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
 6481 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
 6541 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
 6601 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
 6661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
 6721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
 6781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
 6841 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
 6901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
 6961 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
 7021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
 7081 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
 7141 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
 7201 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
 7261 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
 7321 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
 7381 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
 7441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
 7501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
 7561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
 7621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
 7681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
 7741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
 7801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
 7861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
 7921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
 7981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
 8041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
 8101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
 8161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
 8221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
 8281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
 8341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
 8401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
 8461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
 8521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
 8581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
 8641 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
 8701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
 8761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
 8821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
 8881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
 8941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
 9001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
 9061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
 9121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt

//