LOCUS       V014060                12143 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014060
VERSION     V014060
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12143)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..18)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_RNA        596..671
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        711..815
                     /label="AmpR promoter"
     CDS             816..1673
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1847..2435
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2723..2744
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2759..2789
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     CDS             3587..4300
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"
     CDS             5616..6176
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             6219..6689
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6690..7334
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             7524..11624
                     /label="Cas9"
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
ORIGIN
        1 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
       61 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
      121 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ccactatcca actacactca
      181 taatgcttaa agaaaattgg cattatttta ccaatttttt taaaacaaaa aaagaataaa
      241 atattttctt attatttagc taaaattgta ctaaatttta taccaaaaaa aatatatata
      301 aatatatgta atatataata tatatatata tatatgtata tatgtatgta tgtatatatg
      361 tatgtatgta tatatgtatg tatgtatata tgtatgtatg tatgtatagt ttatatttat
      421 ttggcaagtt tttctgtaaa aatgtttaat taaaaaaaaa aatatataac atgccaatat
      481 gcactattca ttttatggag gcaattttat gagtaaccat ttttactttt tttttacata
      541 tgcgcatact tcgagttata caatgttatt ggagacgcgt agaggcctcg tctcagtttt
      601 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      661 cgagtcggtg ctttttttgg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      721 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      781 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      841 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      901 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      961 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1021 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1081 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1141 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1201 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1261 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1321 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1381 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1441 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1501 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1561 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1621 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1681 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1741 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1981 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2221 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2461 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2521 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2581 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2641 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2701 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2761 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2821 caggaaacag ctatgaccat gattacgcca agcttcaaca tatgctgata accaaccagg
     2881 agtcttaata caagtatatg aaggagaaag agctttaaca aaagataata atttattagg
     2941 taaatttcat ttagatggta tcccacctgc tcctagaaaa gtaccacaaa ttgaagttac
     3001 attcgatatt gatgctaatg gtattttaaa tgttacagct gttgaaaaaa gtactggtaa
     3061 acaaaaccat attacaatta caaatgataa aggtagatta tctcctgaag aaattgatag
     3121 aatggttaat gatgctgaaa aatataaagc agaagatgaa gaaaataaaa aaagaattga
     3181 agccagaaat agtcttgaaa attattgcta tggtgttaaa agttcattag aagatcaaaa
     3241 aattaaagaa aaattacaac ctaatgaagt tgaaacatgt atgaaatctg ttacttctat
     3301 tcttgaatgg ttagaaaaaa atcaattagc tggaaaggat gaatatgaag ctaaacaaaa
     3361 agaagctgaa gctgtatgct ctccaattat gtctaaaata tatcaagatg ctggtgcagc
     3421 tgctggtggt atgccaggtg gaatgccagg tggaatgcca ggaggaatgc caggaggaat
     3481 gccaggagga atgaacttcc caggtggaat gccaggagga atgggagcac ccgctggagc
     3541 cccagcgggc tcgggaccaa ctgttgaaga agttgatggt tcgggaatga gtaaaggaga
     3601 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     3661 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     3721 atttatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg
     3781 ttatggtgtt caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa
     3841 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
     3901 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     3961 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     4021 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt
     4081 caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa
     4141 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc
     4201 tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
     4261 agctgctggg attacacatg gcatggatga actatacaaa taaataaatt aatttttttt
     4321 tttttttttt ttttatattg ttgttctgta gttcttcttt tgtgaatagt atttttatat
     4381 atgtactgtt aattttaaaa tgtggaacgc cacgaacata ttttctcgaa caaaatatat
     4441 attttttgtg tcgaaatata taaatttgtg tcgaatatat aaatttgtgt acttcctttt
     4501 tgttgtatgc atgaaaaaaa ggaataacat caaagggggt cgagttatta tgctctctag
     4561 accctaaaac aaacttgtac atcaccacta ctttaatcat acgtaatatc actactttaa
     4621 gcatacgtaa tatcactact ttaatcatac gtaatatcat tagatgtaat atcatcagat
     4681 gtaatatcat cagatgtaat atcatcagat gtaatatcat taaatgtgaa agcgtgttag
     4741 cacttggtat cggtttagct ttccagtcat gctggcaatc acatgctaat atgtgaggtg
     4801 tgcccttttt aactacactc aaaataacat gatttaatta acattggcga gcagcatatc
     4861 acttttgatg cactagctct gaaatgtgta caagttttca tttgctaaag ttacaaaaaa
     4921 agggggagtt tttatttgag gcaagtgcgt taaaatggaa aattaaaaaa atgaaaaaat
     4981 gaaaaaaaat gaaaaattaa aaaaaatgaa aaaatgaaag aatggcagaa tggtgaaaga
     5041 gcgggaggtg gcgaattcct taagtttaca atttaatatt catactttaa gtattttttg
     5101 tagtatccta gatattgtgc tttaaatgct cacccctcaa agcaccagta atattttcat
     5161 ccactgaaat accattaaat tttcaaaaaa atactatgca tataatgtta tacatataaa
     5221 cataaaacgc catgtaaatc aaaaaatata taaaaatatg tataaaaata aatatgcact
     5281 aaatataagc taattatgca taaaaattaa agtgcccttt attaactagt cgtaattatt
     5341 tatatttcta tgttataaaa aaatcctcat ataataatat aattaatata tgtaatgttt
     5401 tttttatttt ataattttaa tataaaataa tatgtaaatt aattcaaaaa ataaatataa
     5461 ttgttgtgaa acaaaaaacg taattttttc atttgccttc aaaatttaaa tttattttaa
     5521 tatttcctaa aatatatata ctttgtgtat aaatatataa aaatatatat ttgcttataa
     5581 ataaataaaa aattttataa aacatagggg gatctatggt tggttcgcta aactgcatcg
     5641 tcgctgtgtc ccagaacatg ggcatcggca agaacgggga cctgccctgg ccaccgctca
     5701 ggaacgaatt tagatatttc cagagaatga ccacaacctc ttcagtagaa ggtaaacaga
     5761 atctggtgat tatgggtaag aagacctggt tctccattcc tgagaagaat cgacctttaa
     5821 agggtagaat taatttagtt ctcagcagag aactcaagga acctccacaa ggagctcatt
     5881 ttctttccag aagtctagat gatgccttaa aacttactga acaaccagaa ttagcaaata
     5941 aagtagacat ggtctggata gttggtggca gttctgttta taaggaagcc atgaatcacc
     6001 caggccatct taaactattt gtgacaagga tcatgcaaga ctttgaaagt gacacgtttt
     6061 ttccagaaat tgatttggag aaatataaac ttctgccaga atacccaggt gttctctctg
     6121 atgtccagga ggagaaaggc attaagtaca aatttgaagt atatgagaag aatgatgcaa
     6181 gcggaggagg tggatctggt ggaggtggaa gtgcaagcgt gacaggggga atggctagca
     6241 agtgggatca gaagggtatg gacattgcct atgaggaggc ggccttaggt tacaaagagg
     6301 gtggtgttcc tattggcgga tgtcttatca ataacaaaga cggaagtgtt ctcggtcgtg
     6361 gtcacaacat gagatttcaa aagggatccg ccacactaca tggtgagatc tccactttgg
     6421 aaaactgtgg gagattagag ggcaaagtgt acaaagatac cactttgtat acgacgctgt
     6481 ctccatgcga catgtgtaca ggtgccatca tcatgtatgg tattccacgc tgtgttgtcg
     6541 gtgagaacgt taatttcaaa agtaagggcg agaaatattt acaaactaga ggtcacgagg
     6601 ttgttgttgt tgacgatgag aggtgtaaaa agatcatgaa acaatttatc gatgaaagac
     6661 ctcaggattg gtttgaagat attggtgagg cttcggaacc atttaagaac gtctacttgc
     6721 tacctcaaac aaaccaattg ctgggtttgt acaccatcat cagaaataag aatacaacta
     6781 gacctgattt cattttctac tccgatagaa tcatcagatt gttggttgaa gaaggtttga
     6841 accatctacc tgtgcaaaag caaattgtgg aaactgacac caacgaaaac ttcgaaggtg
     6901 tctcattcat gggtaaaatc tgtggtgttt ccattgtcag agctggtgaa tcgatggagc
     6961 aaggattaag agactgttgt aggtctgtgc gtatcggtaa aattttaatt caaagggacg
     7021 aggagactgc tttaccaaag ttattctacg aaaaattacc agaggatata tctgaaaggt
     7081 atgtcttcct attagaccca atgctggcca ccggtggtag tgctatcatg gctacagaag
     7141 tcttgattaa gagaggtgtt aagccagaga gaatttactt cttaaaccta atctgtagta
     7201 aggaagggat tgaaaaatac catgccgcct tcccagaggt cagaattgtt actggtgccc
     7261 tcgacagagg tctagatgaa aacaagtatc tagttccagg gttgggtgac tttggtgaca
     7321 gatactactg tgttggttcg ggagagggca gaggatccct gctaacatgc ggtgatgtcg
     7381 aggagaatcc tggcccagaa tcgatggact ataaggacca cgacggagac tacaaggatc
     7441 atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg
     7501 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca
     7561 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     7621 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     7681 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     7741 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     7801 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     7861 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     7921 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     7981 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     8041 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     8101 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     8161 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     8221 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     8281 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     8341 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     8401 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     8461 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     8521 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     8581 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     8641 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     8701 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     8761 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     8821 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     8881 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     8941 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     9001 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     9061 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     9121 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     9181 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     9241 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     9301 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     9361 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     9421 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     9481 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     9541 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     9601 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     9661 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     9721 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     9781 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     9841 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     9901 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     9961 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
    10021 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
    10081 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
    10141 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
    10201 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
    10261 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
    10321 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
    10381 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
    10441 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
    10501 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
    10561 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
    10621 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
    10681 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
    10741 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
    10801 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
    10861 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
    10921 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
    10981 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
    11041 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
    11101 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
    11161 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
    11221 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
    11281 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
    11341 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
    11401 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
    11461 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
    11521 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
    11581 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca
    11641 cgaaaaaggc cggccaggca aaaaagaaaa agtaagaatt accggtgatc ccgtttttct
    11701 tacttatata tttataccaa ttgattgtat ttataactgt aaaaatgtgt atgttgtgtg
    11761 catatttttt tttgtgcatg cacatgcatg taaatagcta aaattatgaa cattttattt
    11821 tttgttcaga aaaaaaaaac tttacacaca taaaatggct agtatgaata gccatatttt
    11881 atataaatta aatcctatga atttatgacc atattaaaaa tttagatatt tatggaacat
    11941 aatatgtttg aaacaataag acaaaattat tattattatt attattttta ctgttataat
    12001 tatgttgtct cttcaatgat tcataaatag ttggacttga tttttaaaat gtttataata
    12061 tgattagcat agttaaataa aaaaagttga aaaattaaaa aaaaacatat aaacacaaat
    12121 gatgtttttt ccttcaattt cgg
//
