LOCUS       V015969                 6176 bp    DNA     circular SYN 14-APR-2019
DEFINITION  Exported.
ACCESSION   V015969
VERSION     V015969
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6176)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz-Castro S, Johnson AD, Bennett
            RJ.
  TITLE     Genetic Modification of Closely Related Candida Species
  JOURNAL   Front Microbiol 10, 357 (2019)
   PUBMED   30941104
REFERENCE   2  (bases 1 to 6176)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2019) Departmento de Ingenieria Genetica, Centro
            de Investigacion y de Estudios Avanzados del Instituto Politecnico
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico
REFERENCE   3  (bases 1 to 6176)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front
            Microbiol 10, 357 (2019)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-JAN-2019) Departmento de Ingenieria Genetica, Centro de
            Investigacion y de Estudios Avanzados del Instituto Politecnico
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico"
FEATURES             Location/Qualifiers
     source          1..6176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label="lac operator"
                     /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)."
     primer_bind     205..221
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        239..257
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    337..1322
                     /gene="LEU2"
                     /label="contains promoter and 5' UTR"
                     /note="contains promoter and 5' UTR"
     CDS             1323..2441
                     /gene="LEU2"
                     /label="3-isopropylmalate dehydrogenase"
                     /note="3-isopropylmalate dehydrogenase from Candida
                     albicans. Accession#: P87186"
     misc_feature    2445..2993
                     /gene="LEU2"
                     /label="contains 3' UTR and terminator"
                     /note="contains 3' UTR and terminator"
     promoter        complement(3064..3082)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3089..3105)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             3243..3542
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             3894..4685
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             4895..5266
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     rep_origin      5407..5995
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttttat ggccgtcaat aatgaaattg
      361 gtgtcatcca acctttgaaa gaaattggta aaatatgtcg tgaaaataaa gtgtttttcc
      421 acactgatgc tgctcaggcg tatggtaaga ttcctattga tgttaacgag atgaatattg
      481 acttgttgtc tatttcttcc cataaaatct atggtccaaa aggtattggt gcctgttatg
      541 ttagaagaag accaagagtg agattggatc caatcattac tggtggtggt caagaaagag
      601 gtttgcgttc aggtacttta gctcctccat tagttgctgg gtttggtgaa gcagcaagat
      661 taatgaagca agaatcgctg tttgacaaaa gacacattga gaaattatcc tctaagttga
      721 aaaatggatt gttgtcaatt ccatctaccc aattcaacgg ttgtaacgac gccaaatctc
      781 aataccctgg ttgcgtgaat gtatcttttg cttacattga aggtgaatcg ttgttaatgg
      841 ccttgaaaga tattgctttg agttctgggt cagcatgtac ttcggcatct ttggaaccat
      901 catatgtttt gcatgcattg ggagctgatg atgccttagc tcattcttca attagatttg
      961 gtattggtag atttacaact gaagccgaag tcgactatgt cattcaagca attaatgaaa
     1021 gagttgactt tttaagaaaa atgtccccgt tatgggaaat ggttcaagaa ggtattgatt
     1081 tgaatacaat tgaatggagt ggtcattagg tagtttttgt tggtgtattt atcatgtata
     1141 gaaagagatt actttgaata attttttttt tgcctgtacc ggagcgagca tgaggttgtg
     1201 attttgctat tccggcgctc cttctggggt gccctttttt caacaaatga attttttttt
     1261 tttcactcca gaaatactcc ccttttctta ttcctttctt ttaaaaccaa tatccaaaaa
     1321 acatgtctgt taaaaccaaa accattactg tcttgcctgg tgaccatgtc ggtaccgaaa
     1381 ttgtcaatga agcaatcaaa gtcttgaaag caattgaagc tgccactccc taccaaaaaa
     1441 tccaatttga tttcaaacac cacttgattg gtggtgctgc cattgatgcc actggcgtcc
     1501 cgcttccaga tgatgctctt gaaagtgcta aatcatccga tgcagttttg ttgggtgctg
     1561 ttggtggacc caaatggggt actggcactg ttcgtcctga acaaggatta ttgaaaatcc
     1621 gtaaagaatt gaacctttat gccaacatta gaccatgtaa ttttgccagt gactcgttat
     1681 tagaattgtc tccattaaaa gctgaagtgg tcaaaggaac caatttaatt attgttcgtg
     1741 aattagttgg tggtatatat tttggtgaac gtcaagaaca agaagaaagt gaagacaaaa
     1801 aaacagcttg ggatacagaa aagtatactg ttgacgaagt tactcgtatc actcgtatgg
     1861 cagcatttat ggctttgcag cacaatccac cattgccaat atggtctttg gacaaagcta
     1921 atgttttggc atcgtcgaga ttgtggagaa agaccgttga taaagtgatt tctgaagaat
     1981 ttccagcttt gtctgtgcag catcaattga ttgattcggc tgcaatgatt ttgatccaaa
     2041 acccaaccaa attgaacggt atcatcatta catctaacat gtttggtgat atcatctcag
     2101 atgaggcatc cgttatccca ggatctttgg gtttattgcc atctgcttca ttggcttcat
     2161 tgccagatac caacactgct tttggtcttt atgagccttg ccatgggtct gctccagatt
     2221 taccagccaa caaagtgaat ccaattgcta caatcttgtc tgccgcttcc atgttgagat
     2281 tatccttgga ttgtgttaag gaagctgaag cattggaaga agctgtcaaa caggttttgg
     2341 atagcggtat cagaacagca gatttaagag gtactagctc tactacagaa gtcggtgacg
     2401 caatagccga aacagtcact aaaattttaa aacaaaatac ataaacagta tatacagtag
     2461 ttagcattta aatttcaaat actccaaagt ctacagattt ctctaaatat gcacacaact
     2521 gagttaccga atcttcactc aataaaggtt gcttatttgc aaaatttttt tgcctggtag
     2581 atgtaatggt ggcctcgctt gattcggatt gacttggctc tcgtatatga tttggctctt
     2641 cattcggaga gaaaaactct tcatcctcac tactgctcgt caatgttata gcgctggtat
     2701 tgtgaggcaa cagaggtttc gacaacttgg gcacgggctc gtttccaaat tcgttgtcaa
     2761 taaactgact gaattcattt gtcatttccg aaaccgtgat agaaacagct gactcgtcac
     2821 gtgtgatttg ttggggtttt agtggtcgtg gtggtggcgg tggtgcaaac atcgttttcg
     2881 agaaagctct attggctgat tttgttgttc tgataatctt tgggatagag tctggtctct
     2941 ttgttgggta gcgatctatt tttttagttt tgaataaatt ggaaacatca acaaagggcg
     3001 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggcccaatt
     3061 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     3121 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3181 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac
     3241 ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     3301 agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc
     3361 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     3421 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     3481 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     3541 tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga
     3601 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     3661 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     3721 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     3781 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct
     3841 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3901 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3961 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4021 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4081 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4141 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4201 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4261 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4321 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4381 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4441 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4501 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4561 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4621 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4681 tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     4741 gcggtatttc acaccgcata caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4861 aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc
     4921 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     4981 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     5041 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     5101 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     5161 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     5221 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctaaa
     5281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5521 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6001 cagcaacgcg gcctttttac ggttcctggg cttttgctgg ccttttgctc acatgttctt
     6061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
