LOCUS       V015758                 6763 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V015758
VERSION     V015758
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F,
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     A new toolkit for gene tagging in Candida albicans containing
            recyclable markers
  JOURNAL   PLoS ONE (2019) In press
REFERENCE   2  (bases 1 to 6763)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F,
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2019) Instituto de Biologia Funcional y Genomica
            (IBFG), Consejo Superior de Investigaciones Cientificas, Zacarias
            Gonzalez 2, Salamanca, Salamanca 37007, Spain
REFERENCE   3  (bases 1 to 6763)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 13
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE
            (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-MAR-2019) Instituto de Biologia Funcional y Genomica (IBFG),
            Consejo Superior de Investigaciones Cientificas, Zacarias Gonzalez
            2, Salamanca, Salamanca 37007, Spain"
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..69
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    83..116
                     /label="loxP"
                     /note="loxP"
     CDS             546..1355
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      1500..2831
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(2859..3262,3427..4054)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59752.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3263..3426
                     /note="modified TUB2 intron sequence"
     3'UTR           4067..4197
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4198..4257
                     /label="derived from Saccharomyces cerevisiae CYC1"
                     /note="derived from Saccharomyces cerevisiae CYC1"
                     /regulatory_class="terminator"
     protein_bind    complement(4270..4303)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4401..4419)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(4677..5265)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5439..6296)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6297..6401)
                     /label="AmpR promoter"
     promoter        6747..6763
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggcggcc gctctagaac
       61 tagtggatcc agatcttgat ccataacttc gtatagcata cattatacga agttatagat
      121 ccagtactaa taggaattga tttggatggt ataaacggaa acaaaaaaaa gagctggtac
      181 tactttcttt aaaattattt tattatttga ttttatttaa tagtatatat tatattttga
      241 acgtagatta ttttgttgaa agttgctgta gtgccattga ttcgtaacac taattctgta
      301 ttagtcattc ctcttgtttg atagtatcca aaaaaacggc tatttttttg caatcttatt
      361 tcctgcatat tatacagata acataatgaa agaaaaaatc tttttttttg ttcttcaatg
      421 atgatttcaa ccattctttt aaacattgat caattcctga gcaacaaccc catacacact
      481 ggtttatata tcgccccttt tacagttgaa gaaagaaata gaaatagaaa tagcaaacaa
      541 aagatatgac agtcaacact aagacctata gtgagagagc agaaactcat gcctcaccag
      601 tagcacaacg attatttcga ttaatggaac tgaagaaaac caatttatgt gcatcaattg
      661 atgttgatac cactaaggaa ttccttgaat taattgataa attgggtcct tatgtatgct
      721 taatcaagac tcatattgat ataatcaatg atttttccta tgaatccact attgaaccat
      781 tattagaact ttcacgtaaa catcaattta tgatttttga agatagaaaa tttgctgata
      841 ttggtaatac cgtgaagaaa caatatattg gtggagttta taaaattagt agttgggcag
      901 atattactaa tgctcatggt gtcactggga atggagtagt tgaaggatta aaacagggag
      961 ctaaagaaac caccaccaac caagagccaa gagggttatt gatgttagct gaattatcat
     1021 cagtgggatc attagcatat ggagaatatt ctcaaaaaac tgttgaaatt gctaaatccg
     1081 ataaggaatt tgttattgga tttattgccc aacgtgatat gggtggacaa gaagaaggat
     1141 ttgattggct tattatgaca cctggagttg gattagatga taaaggtgat ggattaggac
     1201 aacaatatag aactgttgat gaagttgtta gcactggaac tgatattatc attgttggta
     1261 gaggattgtt tggtaaagga agagatccag atattgaagg taaaaggtat agagatgctg
     1321 gttggaatgc ttatttgaaa aaaactggcc aattataaat gtgaaggggg agattttcac
     1381 tttattagat ttgtatatat gtataataaa taaataaata agttaaataa ataattaaat
     1441 aagggtggta attattacta tttacaatca aaggtggtcc ttctagaaga tctgctagca
     1501 attgtctatt ccaagcctgt ggcttttttt ggtgattcac aatacataaa ttggcaaata
     1561 cctcttctat agactttaac ctttgatgaa atggtatgga taaaccactt ttgtggtcag
     1621 ggtatatata tttttattcc aatctggcct ttataaattc aaatttccaa ggggactctg
     1681 ctaaaccttc tgatataact ataactctaa taatggctgt aaaaagtgtg aaaaaaagat
     1741 gatattcaca aaaaagataa aggggatttc atattgggta tctagattta aacatcgtat
     1801 ttcaacaatg aaactagaat caatacgaaa tccagcgcac aactgtgttc attcccttgg
     1861 gaattatact gagtaatggg aagaatgtgc aaaaaatacc cgtgtcttat ataatttcca
     1921 tttgtgtagg gcttaaaagt ggacaacttc aggttacgaa accacgaaaa tgtctaaacg
     1981 tttgatcaaa ttcgtaactc atggaaaatc acagcaacaa attaaggatg gatatatccg
     2041 gtacttacaa ataaaaacac taaagaagta taaattttga actgaaattc tatagaaaat
     2101 cttttttttt gggtaaaatt atgcgattgt ggctcatagt aacgtaatca tacagttcaa
     2161 caaacatcat tttgtcaaag gatttctaat cccattcttt ttctacttgg cttttgagta
     2221 tatatatata catcctttgt tcatcatcat gttcgattga atttttttct tggttgttat
     2281 tagattgtct acttgagttt ttcttggtgc aaattgtggt gtttttctaa caaatttttc
     2341 aacatttaaa gtatttctgg aacttttcta caaggaacga catagatcca ttaatgcgcc
     2401 cctctaaaac aatacccata atgtcttctt aagtatgcat ctgcttataa atgtttctgg
     2461 atttacttta gttagaatgt cgacacgaca atgaactatt gctgtggatc acgtgcgatc
     2521 actgaattat gagtttcttt ttatttttct ttttgggctt ttttttgcaa tttttttttt
     2581 ccacctttgc taattgcgta gtcacatata atgtatacaa tttttgcaac tctgacacta
     2641 taattgatcc aataataaag tatataatag atcatcaagt atacgtaatc tccccaaaaa
     2701 aaaattgcat aaaataaaat ctataaacct tttttagatt tttagttgtt ccttttttct
     2761 ttctattctt caacttccaa gtgttgtcac tttctccctg ttaactatat ttaaaagtaa
     2821 ataccctccc ccccgggtat actcattaat caataattat gtctaacttg ttgaccgttc
     2881 accaaaactt gccagctttg ccagttgatg ctacctctga tgaagttaga aaaaacttga
     2941 tggatatgtt tagagataga caagccttct ctgaacacac ctggaaaatg ttgttgtctg
     3001 tttgtagatc ctgggctgct tggtgtaaat tgaacaacag aaaatggttc ccagccgaac
     3061 cagaagatgt tagagattac ttgttgtact tgcaagctag aggtttggct gttaaaacca
     3121 ttcaacaaca cttgggtcaa ttgaacatgt tgcacagaag atccggtttg ccaagaccat
     3181 ctgattctaa cgctgtttct ttggttatga gaagaattag aaaagaaaac gttgatgctg
     3241 gtgaaagagc taaacaagcc ttgtaggtat agacactgaa gaaaaaaaaa ttctatcatt
     3301 gttatgttgt gatctttgat ctttagttgt cgggttaaca cctgccaatt ggatcaatac
     3361 atcaatcaat taattctaat cttgaaaaaa aaattcattt tactaacaat tttttctttt
     3421 atttagggct ttcgaaagaa ccgatttcga tcaagttaga tccttgatgg aaaactctga
     3481 tagatgtcaa gatattagaa acttggcttt cttgggtatt gcttacaaca ccttgttgag
     3541 aattgctgaa attgctagaa ttagagttaa agatatttcc agaaccgatg gtggtagaat
     3601 gttgattcac attggtagaa ccaaaacctt ggtttctacc gctggtgttg aaaaagcctt
     3661 gtctttgggt gttaccaaat tggttgaaag atggatttct gtttctggtg ttgctgatga
     3721 tccaaacaac tacttgttct gtagagttag aaaaaacggt gttgctgctc catctgctac
     3781 ctctcaatta tctaccagag ctttggaagg tattttcgaa gctacccaca gattgattta
     3841 cggtgctaaa gatgattctg gtcaaagata cttggcttgg tctggtcact ctgctagagt
     3901 tggtgctgct agagatatgg ctagagccgg cgtttctatt ccagaaatta tgcaagctgg
     3961 tggttggacc aacgttaaca ttgttatgaa ctacattaga aacttggatt ctgaaaccgg
     4021 tgctatggtt agattgttgg aagatggtga ttaatttata cctaggtaag caaatagcta
     4081 aattatatac gaattaatat tatgattaag tgtttacgtg agtgcgatat ttttattact
     4141 atcttataca gttgtatata ctctataaaa tgagttgtct attaattaac gcgatgagtc
     4201 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattaat
     4261 tatccatgga taacttcgta tagcatacat tatacgaagt tattagggat aacagggtaa
     4321 tctgcaggaa aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg
     4381 gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     4441 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4501 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4561 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4621 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4681 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4741 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4801 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4861 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4921 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4981 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5041 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5101 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5161 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5221 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5281 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5341 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5401 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5461 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5521 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5581 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5641 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5701 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5761 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5821 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5881 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5941 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6001 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6061 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6121 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6181 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6241 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6301 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6361 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6421 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6481 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6541 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6601 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6661 attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac
     6721 ataaccttat gtatcataca catacgattt aggtgacact ata
//
