LOCUS       V014019                11498 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014019
VERSION     V014019
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11498)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11498
                     /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             4971..5531
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             5574..6044
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6045..6689
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             6879..10979
                     /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 taattggcga
     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 agctggaaca aaattatata tagcaccgaa ttccttaagt ttacaattta atattcatac
     4441 tttaagtatt ttttgtagta tcctagatat tgtgctttaa atgctcaccc ctcaaagcac
     4501 cagtaatatt ttcatccact gaaataccat taaattttca aaaaaatact atgcatataa
     4561 tgttatacat ataaacataa aacgccatgt aaatcaaaaa atatataaaa atatgtataa
     4621 aaataaatat gcactaaata taagctaatt atgcataaaa attaaagtgc cctttattaa
     4681 ctagtcgtaa ttatttatat ttctatgtta taaaaaaatc ctcatataat aatataatta
     4741 atatatgtaa tgtttttttt attttataat tttaatataa aataatatgt aaattaattc
     4801 aaaaaataaa tataattgtt gtgaaacaaa aaacgtaatt ttttcatttg ccttcaaaat
     4861 ttaaatttat tttaatattt cctaaaatat atatactttg tgtataaata tataaaaata
     4921 tatatttgct tataaataaa taaaaaattt tataaaacat agggggatct atggttggtt
     4981 cgctaaactg catcgtcgct gtgtcccaga acatgggcat cggcaagaac ggggacctgc
     5041 cctggccacc gctcaggaac gaatttagat atttccagag aatgaccaca acctcttcag
     5101 tagaaggtaa acagaatctg gtgattatgg gtaagaagac ctggttctcc attcctgaga
     5161 agaatcgacc tttaaagggt agaattaatt tagttctcag cagagaactc aaggaacctc
     5221 cacaaggagc tcattttctt tccagaagtc tagatgatgc cttaaaactt actgaacaac
     5281 cagaattagc aaataaagta gacatggtct ggatagttgg tggcagttct gtttataagg
     5341 aagccatgaa tcacccaggc catcttaaac tatttgtgac aaggatcatg caagactttg
     5401 aaagtgacac gttttttcca gaaattgatt tggagaaata taaacttctg ccagaatacc
     5461 caggtgttct ctctgatgtc caggaggaga aaggcattaa gtacaaattt gaagtatatg
     5521 agaagaatga tgcaagcgga ggaggtggat ctggtggagg tggaagtgca agcgtgacag
     5581 ggggaatggc tagcaagtgg gatcagaagg gtatggacat tgcctatgag gaggcggcct
     5641 taggttacaa agagggtggt gttcctattg gcggatgtct tatcaataac aaagacggaa
     5701 gtgttctcgg tcgtggtcac aacatgagat ttcaaaaggg atccgccaca ctacatggtg
     5761 agatctccac tttggaaaac tgtgggagat tagagggcaa agtgtacaaa gataccactt
     5821 tgtatacgac gctgtctcca tgcgacatgt gtacaggtgc catcatcatg tatggtattc
     5881 cacgctgtgt tgtcggtgag aacgttaatt tcaaaagtaa gggcgagaaa tatttacaaa
     5941 ctagaggtca cgaggttgtt gttgttgacg atgagaggtg taaaaagatc atgaaacaat
     6001 ttatcgatga aagacctcag gattggtttg aagatattgg tgaggcttcg gaaccattta
     6061 agaacgtcta cttgctacct caaacaaacc aattgctggg tttgtacacc atcatcagaa
     6121 ataagaatac aactagacct gatttcattt tctactccga tagaatcatc agattgttgg
     6181 ttgaagaagg tttgaaccat ctacctgtgc aaaagcaaat tgtggaaact gacaccaacg
     6241 aaaacttcga aggtgtctca ttcatgggta aaatctgtgg tgtttccatt gtcagagctg
     6301 gtgaatcgat ggagcaagga ttaagagact gttgtaggtc tgtgcgtatc ggtaaaattt
     6361 taattcaaag ggacgaggag actgctttac caaagttatt ctacgaaaaa ttaccagagg
     6421 atatatctga aaggtatgtc ttcctattag acccaatgct ggccaccggt ggtagtgcta
     6481 tcatggctac agaagtcttg attaagagag gtgttaagcc agagagaatt tacttcttaa
     6541 acctaatctg tagtaaggaa gggattgaaa aataccatgc cgccttccca gaggtcagaa
     6601 ttgttactgg tgccctcgac agaggtctag atgaaaacaa gtatctagtt ccagggttgg
     6661 gtgactttgg tgacagatac tactgtgttg gttcgggaga gggcagagga tccctgctaa
     6721 catgcggtga tgtcgaggag aatcctggcc cagaatcgat ggactataag gaccacgacg
     6781 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
     6841 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
     6901 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     6961 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     7021 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     7081 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     7141 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     7201 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     7261 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     7321 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     7381 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     7441 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     7501 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     7561 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     7621 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     7681 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     7741 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     7801 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     7861 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     7921 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     7981 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     8041 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     8101 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     8161 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     8221 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     8281 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     8341 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     8401 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     8461 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     8521 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     8581 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     8641 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     8701 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     8761 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     8821 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     8881 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     8941 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     9001 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     9061 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     9121 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     9181 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     9241 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     9301 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     9361 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     9421 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     9481 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     9541 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     9601 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     9661 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     9721 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     9781 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     9841 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     9901 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     9961 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
    10021 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
    10081 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
    10141 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
    10201 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
    10261 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
    10321 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
    10381 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
    10441 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
    10501 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
    10561 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
    10621 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
    10681 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
    10741 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
    10801 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
    10861 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
    10921 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
    10981 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattaccgg
    11041 tgatcccgtt tttcttactt atatatttat accaattgat tgtatttata actgtaaaaa
    11101 tgtgtatgtt gtgtgcatat ttttttttgt gcatgcacat gcatgtaaat agctaaaatt
    11161 atgaacattt tattttttgt tcagaaaaaa aaaactttac acacataaaa tggctagtat
    11221 gaatagccat attttatata aattaaatcc tatgaattta tgaccatatt aaaaatttag
    11281 atatttatgg aacataatat gtttgaaaca ataagacaaa attattatta ttattattat
    11341 ttttactgtt ataattatgt tgtctcttca atgattcata aatagttgga cttgattttt
    11401 aaaatgttta taatatgatt agcatagtta aataaaaaaa gttgaaaaat taaaaaaaaa
    11461 catataaaca caaatgatgt tttttccttc aatttcgg
//
