LOCUS       V015732                 7748 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V015732
VERSION     V015732
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7748)
  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 7748)
  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 7748)
  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..7748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             65..769
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     regulatory      773..1028
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     1038..1054
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1068..1101
                     /label="loxP"
                     /note="loxP"
     CDS             1531..2340
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      2485..3816
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3844..4247,4412..5039)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59764.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4248..4411
                     /note="modified TUB2 intron sequence"
     3'UTR           5052..5182
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5183..5242
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5255..5288)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5386..5404)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5662..6250)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6424..7281)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7282..7386)
                     /label="AmpR promoter"
     promoter        7732..7748
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggtgctg gcgcaggtgc
       61 ttctgtttca aaaggtgaag aagataatat ggctattatt aaagaattta tgagatttaa
      121 agttcatatg gaaggttcag ttaatggtca tgaatttgaa attgaaggtg aaggtgaagg
      181 tagaccatat gaaggtactc aaactgctaa attgaaagtt actaaaggtg gttcattacc
      241 atttgcttgg gatattttgt caccacaatt tatgtatggt tcaaaagctt atgttaaaca
      301 tccagctgat attccagatt atttaaaatt gtcatttcca gaaggtttta aatgggaaag
      361 agttatgaat tttgaagatg gtggtgttgt tactgttact caagattcat cattacaaga
      421 tggtgaattt atttataagg ttaaattgag aggtactaat tttccatcag atggtccagt
      481 tatgcaaaaa aaaactatgg gttgggaagc ttcatcagaa agaatgtatc cagaagatgg
      541 tgctttaaaa ggtgaaatta aacaaagatt gaaattaaaa gatggtggtc attatgatgc
      601 tgaagttaaa actacttata aagctaaaaa accagttcaa ttaccaggtg cttataatgt
      661 taatattaaa ttggatatta cttcacataa tgaagattat actattgttg aacaatatga
      721 aagagctgaa ggtagacatt caactggtgg tatggatgaa ttatataaat aaggcgcgcc
      781 attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca
      841 gttattaccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa aaaaaattgg
      901 aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc cagaaccaag
      961 taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa agatggacgc
     1021 atagatccgg cggccgctct agaactagtg gatccagatc ttgatccata acttcgtata
     1081 gcatacatta tacgaagtta tagatccagt actaatagga attgatttgg atggtataaa
     1141 cggaaacaaa aaaaagagct ggtactactt tctttaaaat tattttatta tttgatttta
     1201 tttaatagta tatattatat tttgaacgta gattattttg ttgaaagttg ctgtagtgcc
     1261 attgattcgt aacactaatt ctgtattagt cattcctctt gtttgatagt atccaaaaaa
     1321 acggctattt ttttgcaatc ttatttcctg catattatac agataacata atgaaagaaa
     1381 aaatcttttt ttttgttctt caatgatgat ttcaaccatt cttttaaaca ttgatcaatt
     1441 cctgagcaac aaccccatac acactggttt atatatcgcc ccttttacag ttgaagaaag
     1501 aaatagaaat agaaatagca aacaaaagat atgacagtca acactaagac ctatagtgag
     1561 agagcagaaa ctcatgcctc accagtagca caacgattat ttcgattaat ggaactgaag
     1621 aaaaccaatt tatgtgcatc aattgatgtt gataccacta aggaattcct tgaattaatt
     1681 gataaattgg gtccttatgt atgcttaatc aagactcata ttgatataat caatgatttt
     1741 tcctatgaat ccactattga accattatta gaactttcac gtaaacatca atttatgatt
     1801 tttgaagata gaaaatttgc tgatattggt aataccgtga agaaacaata tattggtgga
     1861 gtttataaaa ttagtagttg ggcagatatt actaatgctc atggtgtcac tgggaatgga
     1921 gtagttgaag gattaaaaca gggagctaaa gaaaccacca ccaaccaaga gccaagaggg
     1981 ttattgatgt tagctgaatt atcatcagtg ggatcattag catatggaga atattctcaa
     2041 aaaactgttg aaattgctaa atccgataag gaatttgtta ttggatttat tgcccaacgt
     2101 gatatgggtg gacaagaaga aggatttgat tggcttatta tgacacctgg agttggatta
     2161 gatgataaag gtgatggatt aggacaacaa tatagaactg ttgatgaagt tgttagcact
     2221 ggaactgata ttatcattgt tggtagagga ttgtttggta aaggaagaga tccagatatt
     2281 gaaggtaaaa ggtatagaga tgctggttgg aatgcttatt tgaaaaaaac tggccaatta
     2341 taaatgtgaa gggggagatt ttcactttat tagatttgta tatatgtata ataaataaat
     2401 aaataagtta aataaataat taaataaggg tggtaattat tactatttac aatcaaaggt
     2461 ggtccttcta gaagatctgc tagcaattgt ctattccaag cctgtggctt tttttggtga
     2521 ttcacaatac ataaattggc aaatacctct tctatagact ttaacctttg atgaaatggt
     2581 atggataaac cacttttgtg gtcagggtat atatattttt attccaatct ggcctttata
     2641 aattcaaatt tccaagggga ctctgctaaa ccttctgata taactataac tctaataatg
     2701 gctgtaaaaa gtgtgaaaaa aagatgatat tcacaaaaaa gataaagggg atttcatatt
     2761 gggtatctag atttaaacat cgtatttcaa caatgaaact agaatcaata cgaaatccag
     2821 cgcacaactg tgttcattcc cttgggaatt atactgagta atgggaagaa tgtgcaaaaa
     2881 atacccgtgt cttatataat ttccatttgt gtagggctta aaagtggaca acttcaggtt
     2941 acgaaaccac gaaaatgtct aaacgtttga tcaaattcgt aactcatgga aaatcacagc
     3001 aacaaattaa ggatggatat atccggtact tacaaataaa aacactaaag aagtataaat
     3061 tttgaactga aattctatag aaaatctttt tttttgggta aaattatgcg attgtggctc
     3121 atagtaacgt aatcatacag ttcaacaaac atcattttgt caaaggattt ctaatcccat
     3181 tctttttcta cttggctttt gagtatatat atatacatcc tttgttcatc atcatgttcg
     3241 attgaatttt tttcttggtt gttattagat tgtctacttg agtttttctt ggtgcaaatt
     3301 gtggtgtttt tctaacaaat ttttcaacat ttaaagtatt tctggaactt ttctacaagg
     3361 aacgacatag atccattaat gcgcccctct aaaacaatac ccataatgtc ttcttaagta
     3421 tgcatctgct tataaatgtt tctggattta ctttagttag aatgtcgaca cgacaatgaa
     3481 ctattgctgt ggatcacgtg cgatcactga attatgagtt tctttttatt tttctttttg
     3541 ggcttttttt tgcaattttt tttttccacc tttgctaatt gcgtagtcac atataatgta
     3601 tacaattttt gcaactctga cactataatt gatccaataa taaagtatat aatagatcat
     3661 caagtatacg taatctcccc aaaaaaaaat tgcataaaat aaaatctata aacctttttt
     3721 agatttttag ttgttccttt tttctttcta ttcttcaact tccaagtgtt gtcactttct
     3781 ccctgttaac tatatttaaa agtaaatacc ctcccccccg ggtatactca ttaatcaata
     3841 attatgtcta acttgttgac cgttcaccaa aacttgccag ctttgccagt tgatgctacc
     3901 tctgatgaag ttagaaaaaa cttgatggat atgtttagag atagacaagc cttctctgaa
     3961 cacacctgga aaatgttgtt gtctgtttgt agatcctggg ctgcttggtg taaattgaac
     4021 aacagaaaat ggttcccagc cgaaccagaa gatgttagag attacttgtt gtacttgcaa
     4081 gctagaggtt tggctgttaa aaccattcaa caacacttgg gtcaattgaa catgttgcac
     4141 agaagatccg gtttgccaag accatctgat tctaacgctg tttctttggt tatgagaaga
     4201 attagaaaag aaaacgttga tgctggtgaa agagctaaac aagccttgta ggtatagaca
     4261 ctgaagaaaa aaaaattcta tcattgttat gttgtgatct ttgatcttta gttgtcgggt
     4321 taacacctgc caattggatc aatacatcaa tcaattaatt ctaatcttga aaaaaaaatt
     4381 cattttacta acaatttttt cttttattta gggctttcga aagaaccgat ttcgatcaag
     4441 ttagatcctt gatggaaaac tctgatagat gtcaagatat tagaaacttg gctttcttgg
     4501 gtattgctta caacaccttg ttgagaattg ctgaaattgc tagaattaga gttaaagata
     4561 tttccagaac cgatggtggt agaatgttga ttcacattgg tagaaccaaa accttggttt
     4621 ctaccgctgg tgttgaaaaa gccttgtctt tgggtgttac caaattggtt gaaagatgga
     4681 tttctgtttc tggtgttgct gatgatccaa acaactactt gttctgtaga gttagaaaaa
     4741 acggtgttgc tgctccatct gctacctctc aattatctac cagagctttg gaaggtattt
     4801 tcgaagctac ccacagattg atttacggtg ctaaagatga ttctggtcaa agatacttgg
     4861 cttggtctgg tcactctgct agagttggtg ctgctagaga tatggctaga gccggcgttt
     4921 ctattccaga aattatgcaa gctggtggtt ggaccaacgt taacattgtt atgaactaca
     4981 ttagaaactt ggattctgaa accggtgcta tggttagatt gttggaagat ggtgattaat
     5041 ttatacctag gtaagcaaat agctaaatta tatacgaatt aatattatga ttaagtgttt
     5101 acgtgagtgc gatattttta ttactatctt atacagttgt atatactcta taaaatgagt
     5161 tgtctattaa ttaacgcgat gagtccctat ttattttttt atagttatgt tagtattaag
     5221 aacgttattt atatttcaaa ttaattatcc atggataact tcgtatagca tacattatac
     5281 gaagttatta gggataacag ggtaatctgc aggaaaaacg agctcgaatt catcgatgat
     5341 atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt
     5401 attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg
     5461 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5521 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5581 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5641 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5701 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5761 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5821 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     5881 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5941 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6001 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6061 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6121 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6181 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6241 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6301 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6361 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6421 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6481 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6541 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6601 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6661 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6721 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6781 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6841 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6901 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6961 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7021 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7081 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7141 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7201 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7261 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7321 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7381 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     7441 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     7501 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     7561 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     7621 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc
     7681 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     7741 acactata
//
