LOCUS       V014018                11481 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014018
VERSION     V014018
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11481)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11481
                     /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             4954..5514
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             5557..6027
                     /gene="FCY1"
                     /label="Cytosine deaminase"
                     /note="Cytosine deaminase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: Q12178"
     CDS             6028..6672
                     /gene="FUR1"
                     /label="Uracil phosphoribosyltransferase"
                     /note="Uracil phosphoribosyltransferase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P18562"
     CDS             6862..10962
                     /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 ttaaagctga
     4381 agctggaaca gaattcctta agtttacaat ttaatattca tactttaagt attttttgta
     4441 gtatcctaga tattgtgctt taaatgctca cccctcaaag caccagtaat attttcatcc
     4501 actgaaatac cattaaattt tcaaaaaaat actatgcata taatgttata catataaaca
     4561 taaaacgcca tgtaaatcaa aaaatatata aaaatatgta taaaaataaa tatgcactaa
     4621 atataagcta attatgcata aaaattaaag tgccctttat taactagtcg taattattta
     4681 tatttctatg ttataaaaaa atcctcatat aataatataa ttaatatatg taatgttttt
     4741 tttattttat aattttaata taaaataata tgtaaattaa ttcaaaaaat aaatataatt
     4801 gttgtgaaac aaaaaacgta attttttcat ttgccttcaa aatttaaatt tattttaata
     4861 tttcctaaaa tatatatact ttgtgtataa atatataaaa atatatattt gcttataaat
     4921 aaataaaaaa ttttataaaa cataggggga tctatggttg gttcgctaaa ctgcatcgtc
     4981 gctgtgtccc agaacatggg catcggcaag aacggggacc tgccctggcc accgctcagg
     5041 aacgaattta gatatttcca gagaatgacc acaacctctt cagtagaagg taaacagaat
     5101 ctggtgatta tgggtaagaa gacctggttc tccattcctg agaagaatcg acctttaaag
     5161 ggtagaatta atttagttct cagcagagaa ctcaaggaac ctccacaagg agctcatttt
     5221 ctttccagaa gtctagatga tgccttaaaa cttactgaac aaccagaatt agcaaataaa
     5281 gtagacatgg tctggatagt tggtggcagt tctgtttata aggaagccat gaatcaccca
     5341 ggccatctta aactatttgt gacaaggatc atgcaagact ttgaaagtga cacgtttttt
     5401 ccagaaattg atttggagaa atataaactt ctgccagaat acccaggtgt tctctctgat
     5461 gtccaggagg agaaaggcat taagtacaaa tttgaagtat atgagaagaa tgatgcaagc
     5521 ggaggaggtg gatctggtgg aggtggaagt gcaagcgtga cagggggaat ggctagcaag
     5581 tgggatcaga agggtatgga cattgcctat gaggaggcgg ccttaggtta caaagagggt
     5641 ggtgttccta ttggcggatg tcttatcaat aacaaagacg gaagtgttct cggtcgtggt
     5701 cacaacatga gatttcaaaa gggatccgcc acactacatg gtgagatctc cactttggaa
     5761 aactgtggga gattagaggg caaagtgtac aaagatacca ctttgtatac gacgctgtct
     5821 ccatgcgaca tgtgtacagg tgccatcatc atgtatggta ttccacgctg tgttgtcggt
     5881 gagaacgtta atttcaaaag taagggcgag aaatatttac aaactagagg tcacgaggtt
     5941 gttgttgttg acgatgagag gtgtaaaaag atcatgaaac aatttatcga tgaaagacct
     6001 caggattggt ttgaagatat tggtgaggct tcggaaccat ttaagaacgt ctacttgcta
     6061 cctcaaacaa accaattgct gggtttgtac accatcatca gaaataagaa tacaactaga
     6121 cctgatttca ttttctactc cgatagaatc atcagattgt tggttgaaga aggtttgaac
     6181 catctacctg tgcaaaagca aattgtggaa actgacacca acgaaaactt cgaaggtgtc
     6241 tcattcatgg gtaaaatctg tggtgtttcc attgtcagag ctggtgaatc gatggagcaa
     6301 ggattaagag actgttgtag gtctgtgcgt atcggtaaaa ttttaattca aagggacgag
     6361 gagactgctt taccaaagtt attctacgaa aaattaccag aggatatatc tgaaaggtat
     6421 gtcttcctat tagacccaat gctggccacc ggtggtagtg ctatcatggc tacagaagtc
     6481 ttgattaaga gaggtgttaa gccagagaga atttacttct taaacctaat ctgtagtaag
     6541 gaagggattg aaaaatacca tgccgccttc ccagaggtca gaattgttac tggtgccctc
     6601 gacagaggtc tagatgaaaa caagtatcta gttccagggt tgggtgactt tggtgacaga
     6661 tactactgtg ttggttcggg agagggcaga ggatccctgc taacatgcgg tgatgtcgag
     6721 gagaatcctg gcccagaatc gatggactat aaggaccacg acggagacta caaggatcat
     6781 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
     6841 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     6901 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     6961 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     7021 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     7081 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     7141 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     7201 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     7261 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     7321 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     7381 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     7441 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     7501 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     7561 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     7621 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     7681 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     7741 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     7801 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     7861 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     7921 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     7981 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     8041 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     8101 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     8161 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     8221 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     8281 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     8341 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     8401 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     8461 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     8521 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     8581 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     8641 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     8701 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     8761 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     8821 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     8881 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     8941 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     9001 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     9061 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     9121 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     9181 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     9241 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     9301 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     9361 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     9421 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     9481 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     9541 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     9601 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     9661 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     9721 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     9781 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     9841 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     9901 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     9961 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
    10021 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
    10081 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
    10141 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
    10201 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
    10261 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
    10321 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
    10381 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
    10441 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
    10501 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
    10561 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
    10621 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
    10681 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
    10741 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
    10801 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
    10861 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
    10921 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
    10981 aaaaaggccg gccaggcaaa aaagaaaaag taagaattac cggtgatccc gtttttctta
    11041 cttatatatt tataccaatt gattgtattt ataactgtaa aaatgtgtat gttgtgtgca
    11101 tatttttttt tgtgcatgca catgcatgta aatagctaaa attatgaaca ttttattttt
    11161 tgttcagaaa aaaaaaactt tacacacata aaatggctag tatgaatagc catattttat
    11221 ataaattaaa tcctatgaat ttatgaccat attaaaaatt tagatattta tggaacataa
    11281 tatgtttgaa acaataagac aaaattatta ttattattat tatttttact gttataatta
    11341 tgttgtctct tcaatgattc ataaatagtt ggacttgatt tttaaaatgt ttataatatg
    11401 attagcatag ttaaataaaa aaagttgaaa aattaaaaaa aaacatataa acacaaatga
    11461 tgttttttcc ttcaatttcg g
//
