LOCUS       V004918                10263 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004918
VERSION     V004918
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10263)
  AUTHORS   Shahana S, Childers DS, Ballou ER, Bohovych I, Odds FC, Gow NA,
            Brown AJ.
  TITLE     New Clox Systems for Rapid and Efficient Gene Disruption in Candida
            albicans
  JOURNAL   PLoS ONE 9 (6), E100390 (2014)
   PUBMED   24940603
REFERENCE   2  (bases 1 to 10263)
  AUTHORS   Shahana S, Childers DS, Brown AJP.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAY-2013) Microbiology, School of Medical Sciences,
            Institute of Medical Sciences - Foresterhill, Aberdeen,
            Aberdeenshire AB25 2ZD, United Kingdom
REFERENCE   3  (bases 1 to 10263)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10263)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
            date: "2014"; volume: "9"; issue: "6"; pages: "E100390"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (01-MAY-2013) Microbiology, School of Medical Sciences, Institute of
            Medical Sciences - Foresterhill, Aberdeen, Aberdeenshire AB25 2ZD,
            United Kingdom"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /label="LEU2"
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(1770..2174)
                     /label="LEU2 promoter"
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3097..3115
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3148..3164
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    3178..3211
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     CDS             3641..4450
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      4595..5926
                     /gene="pMET3"
                     /label="MET3 promoter"
                     /note="MET3 promoter"
                     /regulatory_class="promoter"
     gene            4595..5926
                     /gene="pMET3"
                     /label="pMET3"
     CDS             5954..6361
                     /codon_start=1
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="AGZ63437.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQAL"
     protein_bind    complement(7365..7398)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7443..7459)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(7489..7507)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7528..7544)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    7552..7568
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7576..7606)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7621..7642)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7930..8518)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8692..9549)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9550..9654)
                     /label="AmpR promoter"
     misc_feature    9691..10194
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tccgattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3001 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3061 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     3121 ttggagctcc accgcggtgg cggccgctct agaactagtg gatccagatc ttgatccata
     3181 acttcgtata gcatacatta tacgaagtta tagatccagt actaatagga attgatttgg
     3241 atggtataaa cggaaacaaa aaaaagagct ggtactactt tctttaaaat tattttatta
     3301 tttgatttta tttaatagta tatattatat tttgaacgta gattattttg ttgaaagttg
     3361 ctgtagtgcc attgattcgt aacactaatt ctgtattagt cattcctctt gtttgatagt
     3421 atccaaaaaa acggctattt ttttgcaatc ttatttcctg catattatac agataacata
     3481 atgaaagaaa aaatcttttt ttttgttctt caatgatgat ttcaaccatt cttttaaaca
     3541 ttgatcaatt cctgagcaac aaccccatac acactggttt atatatcgcc ccttttacag
     3601 ttgaagaaag aaatagaaat agaaatagca aacaaaagat atgacagtca acactaagac
     3661 ctatagtgag agagcagaaa ctcatgcctc accagtagca caacgattat ttcgattaat
     3721 ggaactgaag aaaaccaatt tatgtgcatc aattgatgtt gataccacta aggaattcct
     3781 tgaattaatt gataaattgg gtccttatgt atgcttaatc aagactcata ttgatataat
     3841 caatgatttt tcctatgaat ccactattga accattatta gaactttcac gtaaacatca
     3901 atttatgatt tttgaagata gaaaatttgc tgatattggt aataccgtga agaaacaata
     3961 tattggtgga gtttataaaa ttagtagttg ggcagatatt actaatgctc atggtgtcac
     4021 tgggaatgga gtagttgaag gattaaaaca gggagctaaa gaaaccacca ccaaccaaga
     4081 gccaagaggg ttattgatgt tagctgaatt atcatcagtg ggatcattag catatggaga
     4141 atattctcaa aaaactgttg aaattgctaa atccgataag gaatttgtta ttggatttat
     4201 tgcccaacgt gatatgggtg gacaagaaga aggatttgat tggcttatta tgacacctgg
     4261 agttggatta gatgataaag gtgatggatt aggacaacaa tatagaactg ttgatgaagt
     4321 tgttagcact ggaactgata ttatcattgt tggtagagga ttgtttggta aaggaagaga
     4381 tccagatatt gaaggtaaaa ggtatagaga tgctggttgg aatgcttatt tgaaaaaaac
     4441 tggccaatta taaatgtgaa gggggagatt ttcactttat tagatttgta tatatgtata
     4501 ataaataaat aaataagtta aataaataat taaataaggg tggtaattat tactatttac
     4561 aatcaaaggt ggtccttcta gaagatctgc tagcaattgt ctattccaag cctgtggctt
     4621 tttttggtga ttcacaatac ataaattggc aaatacctct tctatagact ttaacctttg
     4681 atgaaatggt atggataaac cacttttgtg gtcagggtat atatattttt attccaatct
     4741 ggcctttata aattcaaatt tccaagggga ctctgctaaa ccttctgata taactataac
     4801 tctaataatg gctgtaaaaa gtgtgaaaaa aagatgatat tcacaaaaaa gataaagggg
     4861 atttcatatt gggtatctag atttaaacat cgtatttcaa caatgaaact agaatcaata
     4921 cgaaatccag cgcacaactg tgttcattcc cttgggaatt atactgagta atgggaagaa
     4981 tgtgcaaaaa atacccgtgt cttatataat ttccatttgt gtagggctta aaagtggaca
     5041 acttcaggtt acgaaaccac gaaaatgtct aaacgtttga tcaaattcgt aactcatgga
     5101 aaatcacagc aacaaattaa ggatggatat atccggtact tacaaataaa aacactaaag
     5161 aagtataaat tttgaactga aattctatag aaaatctttt tttttgggta aaattatgcg
     5221 attgtggctc atagtaacgt aatcatacag ttcaacaaac atcattttgt caaaggattt
     5281 ctaatcccat tctttttcta cttggctttt gagtatatat atatacatcc tttgttcatc
     5341 atcatgttcg attgaatttt tttcttggtt gttattagat tgtctacttg agtttttctt
     5401 ggtgcaaatt gtggtgtttt tctaacaaat ttttcaacat ttaaagtatt tctggaactt
     5461 ttctacaagg aacgacatag atccattaat gcgcccctct aaaacaatac ccataatgtc
     5521 ttcttaagta tgcatctgct tataaatgtt tctggattta ctttagttag aatgtcgaca
     5581 cgacaatgaa ctattgctgt ggatcacgtg cgatcactga attatgagtt tctttttatt
     5641 tttctttttg ggcttttttt tgcaattttt tttttccacc tttgctaatt gcgtagtcac
     5701 atataatgta tacaattttt gcaactctga cactataatt gatccaataa taaagtatat
     5761 aatagatcat caagtatacg taatctcccc aaaaaaaaat tgcataaaat aaaatctata
     5821 aacctttttt agatttttag ttgttccttt tttctttcta ttcttcaact tccaagtgtt
     5881 gtcactttct ccctgttaac tatatttaaa agtaaatacc ctcccccccg ggtatactca
     5941 ttaatcaata attatgtcta acttgttgac cgttcaccaa aacttgccag ctttgccagt
     6001 tgatgctacc tctgatgaag ttagaaaaaa cttgatggat atgtttagag atagacaagc
     6061 cttctctgaa cacacctgga aaatgttgtt gtctgtttgt agatcctggg ctgcttggtg
     6121 taaattgaac aacagaaaat ggttcccagc cgaaccagaa gatgttagag attacttgtt
     6181 gtacttgcaa gctagaggtt tggctgttaa aaccattcaa caacacttgg gtcaattgaa
     6241 catgttgcac agaagatccg gtttgccaag accatctgat tctaacgctg tttctttggt
     6301 tatgagaaga attagaaaag aaaacgttga tgctggtgaa agagctaaac aagccttgta
     6361 ggtatagaca ctgaagaaaa aaaaattcta tcattgttat gttgtgatct ttgatcttta
     6421 gttgtcgggt taacacctgc caattggatc aatacatcaa tcaattaatt ctaatcttga
     6481 aaaaaaaatt cattttacta acaatttttt cttttattta gggctttcga aagaaccgat
     6541 ttcgatcaag ttagatcctt gatggaaaac tctgatagat gtcaagatat tagaaacttg
     6601 gctttcttgg gtattgctta caacaccttg ttgagaattg ctgaaattgc tagaattaga
     6661 gttaaagata tttccagaac cgatggtggt agaatgttga ttcacattgg tagaaccaaa
     6721 accttggttt ctaccgctgg tgttgaaaaa gccttgtctt tgggtgttac caaattggtt
     6781 gaaagatgga tttctgtttc tggtgttgct gatgatccaa acaactactt gttctgtaga
     6841 gttagaaaaa acggtgttgc tgctccatct gctacctctc aattatctac cagagctttg
     6901 gaaggtattt tcgaagctac ccacagattg atttacggtg ctaaagatga ttctggtcaa
     6961 agatacttgg cttggtctgg tcactctgct agagttggtg ctgctagaga tatggctaga
     7021 gccggcgttt ctattccaga aattatgcaa gctggtggtt ggaccaacgt taacattgtt
     7081 atgaactaca ttagaaactt ggattctgaa accggtgcta tggttagatt gttggaagat
     7141 ggtgattaat ttatacctag gtaagcaaat agctaaatta tatacgaatt aatattatga
     7201 ttaagtgttt acgtgagtgc gatattttta ttactatctt atacagttgt atatactcta
     7261 taaaatgagt tgtctattaa ttaacgcgat gagtccctat ttattttttt atagttatgt
     7321 tagtattaag aacgttattt atatttcaaa ttaattatcc atggataact tcgtatagca
     7381 tacattatac gaagttatta gggataacag ggtaatctgc aggaattcga tatcaagctt
     7441 atcgataccg tcgacctcga gggggggccc ggtacccagc ttttgttccc tttagtgagg
     7501 gttaattccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7561 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     7621 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7921 gctggcgttt ttccataggc tcggcccccc tgacgagcat cacaaaaatc gacgctcaag
     7981 tcagaggtgg cgaaacccga caggactata aagataccag gcgttccccc ctggaagctc
     8041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8101 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     8161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8341 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     8401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8761 gcccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9061 ccaacgatca aggcgagtta catgatcccc catgttgtga aaaaaagcgg ttagctcctt
     9121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9661 tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta
     9721 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     9781 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     9841 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     9901 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     9961 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
    10021 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
    10081 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
    10141 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct
    10201 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
    10261 gtc
//
