LOCUS       V014017                11479 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014017
VERSION     V014017
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11479)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11479
                     /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             4952..5512
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             5555..6025
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6026..6670
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             6860..10960
                     /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 agctttgttc cttttgttgt agacatataa
     2881 ttgatcattt tattttttta tttattattt tccaaattct ttaaattttt attaccctta
     2941 aaaaatttcc catttttcca tataaccaaa cttagctagt tatggctagt tttgtatgac
     3001 caatattttt taatattcct aagaattttt ttttttaatt tgtttataaa attatggaaa
     3061 atccaagaaa tcacaattat caatttttta gttaatttca ctttttatgt tttcaataat
     3121 tccatcgaaa agacacgaaa tgatatactc ccttgcactt cacaaaagtg cgaaataaaa
     3181 aatgtatagt aaataatata atattaaata cgtagtattt ttaggctttc ctatattaaa
     3241 ttaattattc tcgacatatt tattaatata tagtaagaaa tgtaaatttt ccaacaagca
     3301 tataacaatt actgttcatt caatattgcc atctaaaatg gaatggaata tatttaaagt
     3361 ggcttcatag aaaggggata ttattaaagg tattattttt aacttttttt tttttattta
     3421 atttttaaaa tttatatttt agtgcactat atctattcgt gaaacatcgg agaaaatata
     3481 tatattcata tatttagatg cacgtggatg catataaacg tatttacaca caccctttcg
     3541 tgtgaactct gattattgat tctatgaatt gaatcatatc aagatgtatc tagggagtgt
     3601 caaaaatgtg gaggatttgt tcctgtcgaa gggagaatac aaacaagtat tcctttgttt
     3661 aaaaaaaaat gaaaatagtg aggcaataca aatagatata agctgtgata gcaacattta
     3721 tagtatctta caattatgca gagaaaaata tggaatccat ggtgattatt taacagatag
     3781 ttgtggaaat attgtagaaa atattaaaga gataatagat ggtgatgctt tatatttaaa
     3841 agataataaa gataaatttc attttttaaa tcaaataaaa tcaaataatt taatagtaaa
     3901 tgattatata gtagaaaaaa gaatagggtc aggtggattt ggtatagtat ttcaaggtat
     3961 acatatacaa actaaacaaa aagttgcact aaaatttata cctaaaagta attttttaga
     4021 tgttacagat gtacatagag tctttataga aatacaaaca ttaagaggtt tgatacataa
     4081 aaatataatt aaaatgtatg atgttaatca ttttcaaaat tatgtatgtc taattatgga
     4141 atatgctgtt aatagtgatt taaaaaaata tatacaaaaa aataatggat atttatctga
     4201 aaaagaaaca tattttttat ttttgcaaat tgttaaagga gtatattatt gtcattctaa
     4261 gcatattgta catagagatt taaaattaga aaatatttta cttgatgaaa atatgacatg
     4321 taaaattgca gattttggat tatctgactt tgttaatgtt gatcaaaata ttaaaactga
     4381 agctggaaga attccttaag tttacaattt aatattcata ctttaagtat tttttgtagt
     4441 atcctagata ttgtgcttta aatgctcacc cctcaaagca ccagtaatat tttcatccac
     4501 tgaaatacca ttaaattttc aaaaaaatac tatgcatata atgttataca tataaacata
     4561 aaacgccatg taaatcaaaa aatatataaa aatatgtata aaaataaata tgcactaaat
     4621 ataagctaat tatgcataaa aattaaagtg ccctttatta actagtcgta attatttata
     4681 tttctatgtt ataaaaaaat cctcatataa taatataatt aatatatgta atgttttttt
     4741 tattttataa ttttaatata aaataatatg taaattaatt caaaaaataa atataattgt
     4801 tgtgaaacaa aaaacgtaat tttttcattt gccttcaaaa tttaaattta ttttaatatt
     4861 tcctaaaata tatatacttt gtgtataaat atataaaaat atatatttgc ttataaataa
     4921 ataaaaaatt ttataaaaca tagggggatc tatggttggt tcgctaaact gcatcgtcgc
     4981 tgtgtcccag aacatgggca tcggcaagaa cggggacctg ccctggccac cgctcaggaa
     5041 cgaatttaga tatttccaga gaatgaccac aacctcttca gtagaaggta aacagaatct
     5101 ggtgattatg ggtaagaaga cctggttctc cattcctgag aagaatcgac ctttaaaggg
     5161 tagaattaat ttagttctca gcagagaact caaggaacct ccacaaggag ctcattttct
     5221 ttccagaagt ctagatgatg ccttaaaact tactgaacaa ccagaattag caaataaagt
     5281 agacatggtc tggatagttg gtggcagttc tgtttataag gaagccatga atcacccagg
     5341 ccatcttaaa ctatttgtga caaggatcat gcaagacttt gaaagtgaca cgttttttcc
     5401 agaaattgat ttggagaaat ataaacttct gccagaatac ccaggtgttc tctctgatgt
     5461 ccaggaggag aaaggcatta agtacaaatt tgaagtatat gagaagaatg atgcaagcgg
     5521 aggaggtgga tctggtggag gtggaagtgc aagcgtgaca gggggaatgg ctagcaagtg
     5581 ggatcagaag ggtatggaca ttgcctatga ggaggcggcc ttaggttaca aagagggtgg
     5641 tgttcctatt ggcggatgtc ttatcaataa caaagacgga agtgttctcg gtcgtggtca
     5701 caacatgaga tttcaaaagg gatccgccac actacatggt gagatctcca ctttggaaaa
     5761 ctgtgggaga ttagagggca aagtgtacaa agataccact ttgtatacga cgctgtctcc
     5821 atgcgacatg tgtacaggtg ccatcatcat gtatggtatt ccacgctgtg ttgtcggtga
     5881 gaacgttaat ttcaaaagta agggcgagaa atatttacaa actagaggtc acgaggttgt
     5941 tgttgttgac gatgagaggt gtaaaaagat catgaaacaa tttatcgatg aaagacctca
     6001 ggattggttt gaagatattg gtgaggcttc ggaaccattt aagaacgtct acttgctacc
     6061 tcaaacaaac caattgctgg gtttgtacac catcatcaga aataagaata caactagacc
     6121 tgatttcatt ttctactccg atagaatcat cagattgttg gttgaagaag gtttgaacca
     6181 tctacctgtg caaaagcaaa ttgtggaaac tgacaccaac gaaaacttcg aaggtgtctc
     6241 attcatgggt aaaatctgtg gtgtttccat tgtcagagct ggtgaatcga tggagcaagg
     6301 attaagagac tgttgtaggt ctgtgcgtat cggtaaaatt ttaattcaaa gggacgagga
     6361 gactgcttta ccaaagttat tctacgaaaa attaccagag gatatatctg aaaggtatgt
     6421 cttcctatta gacccaatgc tggccaccgg tggtagtgct atcatggcta cagaagtctt
     6481 gattaagaga ggtgttaagc cagagagaat ttacttctta aacctaatct gtagtaagga
     6541 agggattgaa aaataccatg ccgccttccc agaggtcaga attgttactg gtgccctcga
     6601 cagaggtcta gatgaaaaca agtatctagt tccagggttg ggtgactttg gtgacagata
     6661 ctactgtgtt ggttcgggag agggcagagg atccctgcta acatgcggtg atgtcgagga
     6721 gaatcctggc ccagaatcga tggactataa ggaccacgac ggagactaca aggatcatga
     6781 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     6841 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     6901 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     6961 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     7021 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     7081 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     7141 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     7201 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     7261 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     7321 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     7381 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     7441 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     7501 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     7561 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     7621 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     7681 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     7741 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     7801 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     7861 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     7921 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     7981 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     8041 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     8101 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     8161 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     8221 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     8281 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     8341 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     8401 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     8461 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     8521 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     8581 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     8641 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     8701 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     8761 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     8821 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     8881 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     8941 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     9001 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     9061 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     9121 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     9181 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     9241 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     9301 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     9361 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     9421 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     9481 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     9541 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     9601 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     9661 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     9721 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     9781 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     9841 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     9901 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     9961 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
    10021 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
    10081 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
    10141 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
    10201 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
    10261 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
    10321 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
    10381 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
    10441 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
    10501 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
    10561 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
    10621 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
    10681 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
    10741 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
    10801 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
    10861 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
    10921 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa
    10981 aaaggccggc caggcaaaaa agaaaaagta agaattaccg gtgatcccgt ttttcttact
    11041 tatatattta taccaattga ttgtatttat aactgtaaaa atgtgtatgt tgtgtgcata
    11101 tttttttttg tgcatgcaca tgcatgtaaa tagctaaaat tatgaacatt ttattttttg
    11161 ttcagaaaaa aaaaacttta cacacataaa atggctagta tgaatagcca tattttatat
    11221 aaattaaatc ctatgaattt atgaccatat taaaaattta gatatttatg gaacataata
    11281 tgtttgaaac aataagacaa aattattatt attattatta tttttactgt tataattatg
    11341 ttgtctcttc aatgattcat aaatagttgg acttgatttt taaaatgttt ataatatgat
    11401 tagcatagtt aaataaaaaa agttgaaaaa ttaaaaaaaa acatataaac acaaatgatg
    11461 ttttttcctt caatttcgg
//
