LOCUS       Exported               10060 bp ds-DNA     circular SYN 16-APR-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    62543
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10060)
  AUTHORS   martin
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 16, 2015 from SnapGene 2.7.1
            http://www.snapgene.com
COMMENT     ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..10060
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        625..651
                     /note="T7"
     promoter        659..1468
                     /note="Rib. Prom"
     misc_feature    1475..1684
                     /note="HX1"
     CDS             1690..5961
                     /codon_start=1
                     /note="3xFLAG-NLS-hSpCas9-NLS"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKYS
                     IGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK
                     RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVD
                     EVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDK
                     LFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLI
                     ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL
                     LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA
                     GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGEL
                     HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNF
                     EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKP
                     AFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHD
                     LLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRR
                     YTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVS
                     GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKG
                     QKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINR
                     LSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKL
                     ITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLI
                     REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFV
                     YGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGE
                     TGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWD
                     PKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKG
                     YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKL
                     KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIRE
                     QAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLS
                     QLGGDKRPAATKKAGQAKKKK"
     CDS             5968..6125
                     /codon_start=1
                     /note="gapdh"
                     /translation="PFTTPRQRCRTTCRRSAASSRLCRGTTTSGDTPTAWWTLYATWPR
                     RIVRQGC"
     5'UTR           6126..6635
                     /note="GAPDH IR"
     misc_feature    6702..7100
                     /note="Blasticidin"
     3'UTR           7115..7856
                     /note="GAPDH 3'UTR"
     CDS             9075..9935
                     /codon_start=1
                     /note="AMP"
                     /translation="LPMLNQ*GTYLSDLSISFIHSCLTPRRVDNYDTGGLTIWPQCCND
                     TARPTLTGSRFISNKPASRKGRAQKWSCNFIRLHPVY*LLPGS*SK*FAS**FAQRCCH
                     CYRHRGVTLVVWYGFIQLRFPTIKASYMIPHVVQKSG*LLRSSDRCQK*VGRSVITHGY
                     GSTA*FSYCHAIRKMLFCDW*VLNQVILRIVYAATELLLPGVNTG*YRAT*QNFKSAHH
                     WKTFFGAKTLKDLTAVEIQFDVTHSCTQLIFSIFYFHQRFWVSKNRKAKCRKKGNKGDT
                     EMLNTH"
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tgcaccctgg gatatttgcg cacccacctt tcccgtcgca atcggcgccg tccgaacgcg
      721 gaaatgttcc gagaataata cggctgggct gcgccgcgcg ttccccgcgt gaaccaacta
      781 ttgtgcgatg tgcagcatcc tattgtggtg ctttaccgca caccagtttc cttgcggggc
      841 ttatttggtg tgttggattt ttaatttttt gatgattcgg aggaattttt tgcaattttt
      901 gggtacccct aaaaaaattt tggagtttcc ggggttcgat ccaacttgtg cttttttaac
      961 cgtccatttg tgtctagtac atcatggagc ggtattctct ccggtggccc aggtggcgtc
     1021 ggaaggaatc ggtgtccctc aggatgtttg gtttttcgtt tttttttttg cggtcttttg
     1081 tttcgcatcg ccgtatcgtg tgcggcgcga ccttcttttt ggctcattgg tcagtgaggt
     1141 ttataataaa gtggatcatt tgggtttttg gccctttgcg gcgcacccgc ggaatggggg
     1201 agggattttc atataattgt agtgtacaca ttgtgtgtgg gtcgcaaaat gtcatgccgt
     1261 gtgcaatgtt tatttgcgtg ctgtgcgtgt ccttttgtga ttccaccact tgtgtgtatt
     1321 ttaccttatc gtctctccct gccgttggct gtgcgtgaat tgtgcttcgc attcgtggga
     1381 aatcaaaagg ggaatcaacg tgttcttttt ttttcagaag gggagtcaaa gcgccgtgtg
     1441 gatgccaagt ctgtgcaaga gttctagtgg atcccccaac gagtttcttc aaaatatgca
     1501 gcggattcac taagaaacat tttcacgcac gaaagcgaaa ttattatgat tgttattata
     1561 atactttttc tttgttgttt tatccactta ttattgttgt gttaaattgt ttttaccttt
     1621 tttcttttcc aacttctttt atgatgtctt ttcttttttt tttttgctct ataagttgtc
     1681 ttgtctagaa tggactataa ggaccacgac ggagactaca aggatcatga tattgattac
     1741 aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc
     1801 ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     1861 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     1921 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     1981 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     2041 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     2101 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     2161 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     2221 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     2281 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     2341 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     2401 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     2461 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     2521 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     2581 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     2641 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     2701 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     2761 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     2821 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     2881 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     2941 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     3001 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     3061 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     3121 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     3181 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     3241 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     3301 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     3361 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     3421 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     3481 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     3541 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     3601 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     3661 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     3721 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     3781 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     3841 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     3901 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     3961 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     4021 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     4081 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     4141 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     4201 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     4261 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     4321 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     4381 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     4441 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     4501 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     4561 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     4621 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     4681 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     4741 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     4801 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     4861 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     4921 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     4981 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     5041 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     5101 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     5161 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     5221 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     5281 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     5341 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     5401 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     5461 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     5521 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     5581 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     5641 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     5701 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     5761 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     5821 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     5881 cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc
     5941 caggcaaaaa agaaaaagta actcgagcca tttacgactc caaggcaacg ctgcagaaca
     6001 acctgccgaa ggagcgccgc ttcttcaaga ttgtgtcgtg gtacgacaac gagtggggat
     6061 actcccaccg cgtggtggac cttgtacgcc acatggcctc gaaggatcgt tcggcaaggt
     6121 tgtaggcgtg gcgatgactt caggtctttc ttttgcgaat agggatctta taatacacga
     6181 tgcgtgtccc gtgatgatcg ttaccggtgc tgccacgatc caattgacac agcgtcaaga
     6241 gcaaaacaat tttacttttc cctttaagga caacaacaaa aaaatatata actttttttt
     6301 cttttttttt ttttttgaaa ttatatttat ggtcatcttt gggaaacaaa aagcagcaat
     6361 ttaatgatgc ggaaggatga gtgaaataat gtttaatcaa tgtacgagga tttggggtat
     6421 tgcaaggaaa atgtagatga tttaattggg tgtgtgatgc agcttgtggt aatttttgct
     6481 cacttccctt tttgccacat ctttttagtt tttctgcttt tctttcccca ttattccact
     6541 tgtctctctt ttcccacgtt tcctgcacga atgcagaaag tgatattttt actttgaaag
     6601 ccatctacca acaacaatta cattgaacag aatttgggat tgcgaattaa ttcctgcagc
     6661 cccgcgtgtg gcctcgaaca ccgagcgacc ctgcagccaa tatggccaag cctttgtctc
     6721 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     6781 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     6841 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     6901 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     6961 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     7021 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     7081 gttatgtgtg ggagggctaa ggggatcgat ccggaacaac ctgccgaagg agcgccgctt
     7141 cttcaagatt gtgtcgtggt acgacaacga gtggggatac tcccaccgcg tggtggacct
     7201 tgtacgccac atggcctcga aggatcgttc ggcaaggttg taggcgtggc gatgacttca
     7261 ggtctttctt ttgcgaatag ggatcttata atacacgatg cgtgtcccgt gatgatcgtt
     7321 accggtgctg ccacgatcca agtgacacag tgtcaagagc aaaacaattt tacttttccc
     7381 tttaaggaca acaacaaaaa aatatataac ttttttttct tttttttttt ttgaaattat
     7441 atttatggtc atctttggga aacaaaaagc agcaatttaa tgatgcggaa ggatgagtga
     7501 aataatgttt aatcaatgta cgaggatttg gggtattgca aggaaaatgt agatgattta
     7561 gttgggtgta tgatgcagct tgtggtaatt ttttacaact ttgcgttatt ggaagatttt
     7621 ggttgttcag tttctttttg tttataactg actgttttaa tttagtgtga tttttttttt
     7681 ttaaggattt aagctgtttt tctttttttt tctttttctt cactgatggt gtttaaggtg
     7741 tagtttagtg tttgtgcatt atgcgtcgta gtgttgttgt tctttggact ttttcgaaca
     7801 atactgggca ttatagccca cctgtgtata tgccgctaga gtatgctagc cgtgtgggta
     7861 cccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     7921 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7981 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8041 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     8101 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     8161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     8761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     9001 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     9061 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     9121 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     9181 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     9241 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     9301 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     9361 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     9421 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     9481 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     9541 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9601 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9661 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9721 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9781 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9841 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9901 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9961 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    10021 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
//
