LOCUS       V014116                12137 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014116
VERSION     V014116
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12137)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12137
                     /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..4294
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             5610..6170
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             6213..6683
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6684..7328
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             7518..11618
                     /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 tcgggaatgg tgagcaaggg
     3601 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     3661 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3721 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3781 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3841 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3901 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3961 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     4021 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     4081 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     4141 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     4201 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     4261 ccactccacc ggcggcatgg acgagctgta caagtagata aattaatttt tttttttttt
     4321 ttttttttat attgttgttc tgtagttctt cttttgtgaa tagtattttt atatatgtac
     4381 tgttaatttt aaaatgtgga acgccacgaa catattttct cgaacaaaat atatattttt
     4441 tgtgtcgaaa tatataaatt tgtgtcgaat atataaattt gtgtacttcc tttttgttgt
     4501 atgcatgaaa aaaaggaata acatcaaagg gggtcgagtt attatgctct ctagacccta
     4561 aaacaaactt gtacatcacc actactttaa tcatacgtaa tatcactact ttaagcatac
     4621 gtaatatcac tactttaatc atacgtaata tcattagatg taatatcatc agatgtaata
     4681 tcatcagatg taatatcatc agatgtaata tcattaaatg tgaaagcgtg ttagcacttg
     4741 gtatcggttt agctttccag tcatgctggc aatcacatgc taatatgtga ggtgtgccct
     4801 ttttaactac actcaaaata acatgattta attaacattg gcgagcagca tatcactttt
     4861 gatgcactag ctctgaaatg tgtacaagtt ttcatttgct aaagttacaa aaaaaggggg
     4921 agtttttatt tgaggcaagt gcgttaaaat ggaaaattaa aaaaatgaaa aaatgaaaaa
     4981 aaatgaaaaa ttaaaaaaaa tgaaaaaatg aaagaatggc agaatggtga aagagcggga
     5041 ggtggcgaat tccttaagtt tacaatttaa tattcatact ttaagtattt tttgtagtat
     5101 cctagatatt gtgctttaaa tgctcacccc tcaaagcacc agtaatattt tcatccactg
     5161 aaataccatt aaattttcaa aaaaatacta tgcatataat gttatacata taaacataaa
     5221 acgccatgta aatcaaaaaa tatataaaaa tatgtataaa aataaatatg cactaaatat
     5281 aagctaatta tgcataaaaa ttaaagtgcc ctttattaac tagtcgtaat tatttatatt
     5341 tctatgttat aaaaaaatcc tcatataata atataattaa tatatgtaat gtttttttta
     5401 ttttataatt ttaatataaa ataatatgta aattaattca aaaaataaat ataattgttg
     5461 tgaaacaaaa aacgtaattt tttcatttgc cttcaaaatt taaatttatt ttaatatttc
     5521 ctaaaatata tatactttgt gtataaatat ataaaaatat atatttgctt ataaataaat
     5581 aaaaaatttt ataaaacata gggggatcta tggttggttc gctaaactgc atcgtcgctg
     5641 tgtcccagaa catgggcatc ggcaagaacg gggacctgcc ctggccaccg ctcaggaacg
     5701 aatttagata tttccagaga atgaccacaa cctcttcagt agaaggtaaa cagaatctgg
     5761 tgattatggg taagaagacc tggttctcca ttcctgagaa gaatcgacct ttaaagggta
     5821 gaattaattt agttctcagc agagaactca aggaacctcc acaaggagct cattttcttt
     5881 ccagaagtct agatgatgcc ttaaaactta ctgaacaacc agaattagca aataaagtag
     5941 acatggtctg gatagttggt ggcagttctg tttataagga agccatgaat cacccaggcc
     6001 atcttaaact atttgtgaca aggatcatgc aagactttga aagtgacacg ttttttccag
     6061 aaattgattt ggagaaatat aaacttctgc cagaataccc aggtgttctc tctgatgtcc
     6121 aggaggagaa aggcattaag tacaaatttg aagtatatga gaagaatgat gcaagcggag
     6181 gaggtggatc tggtggaggt ggaagtgcaa gcgtgacagg gggaatggct agcaagtggg
     6241 atcagaaggg tatggacatt gcctatgagg aggcggcctt aggttacaaa gagggtggtg
     6301 ttcctattgg cggatgtctt atcaataaca aagacggaag tgttctcggt cgtggtcaca
     6361 acatgagatt tcaaaaggga tccgccacac tacatggtga gatctccact ttggaaaact
     6421 gtgggagatt agagggcaaa gtgtacaaag ataccacttt gtatacgacg ctgtctccat
     6481 gcgacatgtg tacaggtgcc atcatcatgt atggtattcc acgctgtgtt gtcggtgaga
     6541 acgttaattt caaaagtaag ggcgagaaat atttacaaac tagaggtcac gaggttgttg
     6601 ttgttgacga tgagaggtgt aaaaagatca tgaaacaatt tatcgatgaa agacctcagg
     6661 attggtttga agatattggt gaggcttcgg aaccatttaa gaacgtctac ttgctacctc
     6721 aaacaaacca attgctgggt ttgtacacca tcatcagaaa taagaataca actagacctg
     6781 atttcatttt ctactccgat agaatcatca gattgttggt tgaagaaggt ttgaaccatc
     6841 tacctgtgca aaagcaaatt gtggaaactg acaccaacga aaacttcgaa ggtgtctcat
     6901 tcatgggtaa aatctgtggt gtttccattg tcagagctgg tgaatcgatg gagcaaggat
     6961 taagagactg ttgtaggtct gtgcgtatcg gtaaaatttt aattcaaagg gacgaggaga
     7021 ctgctttacc aaagttattc tacgaaaaat taccagagga tatatctgaa aggtatgtct
     7081 tcctattaga cccaatgctg gccaccggtg gtagtgctat catggctaca gaagtcttga
     7141 ttaagagagg tgttaagcca gagagaattt acttcttaaa cctaatctgt agtaaggaag
     7201 ggattgaaaa ataccatgcc gccttcccag aggtcagaat tgttactggt gccctcgaca
     7261 gaggtctaga tgaaaacaag tatctagttc cagggttggg tgactttggt gacagatact
     7321 actgtgttgg ttcgggagag ggcagaggat ccctgctaac atgcggtgat gtcgaggaga
     7381 atcctggccc agaatcgatg gactataagg accacgacgg agactacaag gatcatgata
     7441 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
     7501 acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
     7561 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     7621 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     7681 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     7741 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     7801 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     7861 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     7921 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     7981 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     8041 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     8101 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     8161 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     8221 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     8281 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     8341 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     8401 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     8461 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     8521 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     8581 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     8641 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     8701 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     8761 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     8821 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     8881 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     8941 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     9001 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     9061 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     9121 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     9181 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     9241 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     9301 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     9361 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     9421 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     9481 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     9541 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     9601 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     9661 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     9721 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     9781 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     9841 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     9901 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     9961 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
    10021 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
    10081 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
    10141 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
    10201 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
    10261 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
    10321 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
    10381 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
    10441 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
    10501 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
    10561 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
    10621 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
    10681 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
    10741 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
    10801 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
    10861 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
    10921 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
    10981 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
    11041 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
    11101 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
    11161 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
    11221 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
    11281 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
    11341 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
    11401 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
    11461 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
    11521 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
    11581 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
    11641 aggccggcca ggcaaaaaag aaaaagtaag aattaccggt gatcccgttt ttcttactta
    11701 tatatttata ccaattgatt gtatttataa ctgtaaaaat gtgtatgttg tgtgcatatt
    11761 tttttttgtg catgcacatg catgtaaata gctaaaatta tgaacatttt attttttgtt
    11821 cagaaaaaaa aaactttaca cacataaaat ggctagtatg aatagccata ttttatataa
    11881 attaaatcct atgaatttat gaccatatta aaaatttaga tatttatgga acataatatg
    11941 tttgaaacaa taagacaaaa ttattattat tattattatt tttactgtta taattatgtt
    12001 gtctcttcaa tgattcataa atagttggac ttgattttta aaatgtttat aatatgatta
    12061 gcatagttaa ataaaaaaag ttgaaaaatt aaaaaaaaac atataaacac aaatgatgtt
    12121 ttttccttca atttcgg
//
