LOCUS       V014039                11488 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014039
VERSION     V014039
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11488)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11488
                     /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             4961..5521
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             5564..6034
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6035..6679
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             6869..10969
                     /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 ggtgatactt 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 agctggagca aaattatgaa ttccttaagt ttacaattta atattcatac tttaagtatt
     4441 ttttgtagta tcctagatat tgtgctttaa atgctcaccc ctcaaagcac cagtaatatt
     4501 ttcatccact gaaataccat taaattttca aaaaaatact atgcatataa tgttatacat
     4561 ataaacataa aacgccatgt aaatcaaaaa atatataaaa atatgtataa aaataaatat
     4621 gcactaaata taagctaatt atgcataaaa attaaagtgc cctttattaa ctagtcgtaa
     4681 ttatttatat ttctatgtta taaaaaaatc ctcatataat aatataatta atatatgtaa
     4741 tgtttttttt attttataat tttaatataa aataatatgt aaattaattc aaaaaataaa
     4801 tataattgtt gtgaaacaaa aaacgtaatt ttttcatttg ccttcaaaat ttaaatttat
     4861 tttaatattt cctaaaatat atatactttg tgtataaata tataaaaata tatatttgct
     4921 tataaataaa taaaaaattt tataaaacat agggggatct atggttggtt cgctaaactg
     4981 catcgtcgct gtgtcccaga acatgggcat cggcaagaac ggggacctgc cctggccacc
     5041 gctcaggaac gaatttagat atttccagag aatgaccaca acctcttcag tagaaggtaa
     5101 acagaatctg gtgattatgg gtaagaagac ctggttctcc attcctgaga agaatcgacc
     5161 tttaaagggt agaattaatt tagttctcag cagagaactc aaggaacctc cacaaggagc
     5221 tcattttctt tccagaagtc tagatgatgc cttaaaactt actgaacaac cagaattagc
     5281 aaataaagta gacatggtct ggatagttgg tggcagttct gtttataagg aagccatgaa
     5341 tcacccaggc catcttaaac tatttgtgac aaggatcatg caagactttg aaagtgacac
     5401 gttttttcca gaaattgatt tggagaaata taaacttctg ccagaatacc caggtgttct
     5461 ctctgatgtc caggaggaga aaggcattaa gtacaaattt gaagtatatg agaagaatga
     5521 tgcaagcgga ggaggtggat ctggtggagg tggaagtgca agcgtgacag ggggaatggc
     5581 tagcaagtgg gatcagaagg gtatggacat tgcctatgag gaggcggcct taggttacaa
     5641 agagggtggt gttcctattg gcggatgtct tatcaataac aaagacggaa gtgttctcgg
     5701 tcgtggtcac aacatgagat ttcaaaaggg atccgccaca ctacatggtg agatctccac
     5761 tttggaaaac tgtgggagat tagagggcaa agtgtacaaa gataccactt tgtatacgac
     5821 gctgtctcca tgcgacatgt gtacaggtgc catcatcatg tatggtattc cacgctgtgt
     5881 tgtcggtgag aacgttaatt tcaaaagtaa gggcgagaaa tatttacaaa ctagaggtca
     5941 cgaggttgtt gttgttgacg atgagaggtg taaaaagatc atgaaacaat ttatcgatga
     6001 aagacctcag gattggtttg aagatattgg tgaggcttcg gaaccattta agaacgtcta
     6061 cttgctacct caaacaaacc aattgctggg tttgtacacc atcatcagaa ataagaatac
     6121 aactagacct gatttcattt tctactccga tagaatcatc agattgttgg ttgaagaagg
     6181 tttgaaccat ctacctgtgc aaaagcaaat tgtggaaact gacaccaacg aaaacttcga
     6241 aggtgtctca ttcatgggta aaatctgtgg tgtttccatt gtcagagctg gtgaatcgat
     6301 ggagcaagga ttaagagact gttgtaggtc tgtgcgtatc ggtaaaattt taattcaaag
     6361 ggacgaggag actgctttac caaagttatt ctacgaaaaa ttaccagagg atatatctga
     6421 aaggtatgtc ttcctattag acccaatgct ggccaccggt ggtagtgcta tcatggctac
     6481 agaagtcttg attaagagag gtgttaagcc agagagaatt tacttcttaa acctaatctg
     6541 tagtaaggaa gggattgaaa aataccatgc cgccttccca gaggtcagaa ttgttactgg
     6601 tgccctcgac agaggtctag atgaaaacaa gtatctagtt ccagggttgg gtgactttgg
     6661 tgacagatac tactgtgttg gttcgggaga gggcagagga tccctgctaa catgcggtga
     6721 tgtcgaggag aatcctggcc cagaatcgat ggactataag gaccacgacg gagactacaa
     6781 ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa
     6841 ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg
     6901 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     6961 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     7021 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     7081 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     7141 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     7201 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     7261 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     7321 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     7381 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     7441 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     7501 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     7561 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     7621 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     7681 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     7741 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     7801 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     7861 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     7921 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     7981 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     8041 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     8101 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     8161 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     8221 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     8281 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     8341 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     8401 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     8461 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     8521 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     8581 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     8641 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     8701 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     8761 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     8821 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     8881 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     8941 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     9001 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     9061 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     9121 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     9181 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     9241 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     9301 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     9361 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     9421 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     9481 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     9541 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     9601 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     9661 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     9721 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     9781 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     9841 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     9901 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     9961 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
    10021 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
    10081 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
    10141 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
    10201 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
    10261 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
    10321 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
    10381 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
    10441 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
    10501 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
    10561 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
    10621 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
    10681 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
    10741 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
    10801 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
    10861 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
    10921 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
    10981 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattaccgg tgatcccgtt
    11041 tttcttactt atatatttat accaattgat tgtatttata actgtaaaaa tgtgtatgtt
    11101 gtgtgcatat ttttttttgt gcatgcacat gcatgtaaat agctaaaatt atgaacattt
    11161 tattttttgt tcagaaaaaa aaaactttac acacataaaa tggctagtat gaatagccat
    11221 attttatata aattaaatcc tatgaattta tgaccatatt aaaaatttag atatttatgg
    11281 aacataatat gtttgaaaca ataagacaaa attattatta ttattattat ttttactgtt
    11341 ataattatgt tgtctcttca atgattcata aatagttgga cttgattttt aaaatgttta
    11401 taatatgatt agcatagtta aataaaaaaa gttgaaaaat taaaaaaaaa catataaaca
    11461 caaatgatgt tttttccttc aatttcgg
//
