LOCUS       V015620                 9539 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V015620
VERSION     V015620
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9539)
  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 9539)
  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 9539)
  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..9539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(166..465)
                     /label="NEUT5L 3'"
                     /note="NEUT5L 3'"
     primer_bind     736..752
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        759..777
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     817..833
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    847..880
                     /label="loxP"
                     /note="loxP"
     CDS             1310..2119
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      2264..3595
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3623..4026,4191..4818)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59802.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4027..4190
                     /note="modified TUB2 intron sequence"
     3'UTR           4831..4961
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4962..5021
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5034..5067)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5147..5163)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(5174..5423)
                     /label="NEUT5L 5'"
                     /note="NEUT5L 5'"
     promoter        complement(5438..5468)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5483..5504)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5792..6380)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6554..7411)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7412..7516)
                     /label="AmpR promoter"
     misc_feature    7553..8056
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        8312..8592
                     /label="TRP1 promoter"
     CDS             8593..9264
                     /label="TRP1"
                     /note="phosphoribosylanthranilate isomerase, required for
                     tryptophan biosynthesis"
ORIGIN
        1 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
       61 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
      121 agcccccgat ttagagcttg acggggaaag cggccggcta ggccttaacc cactgaattc
      181 tacatcgaac aagagaaacc ttttttttta cttttttcaa acttgaacaa agaaaagaaa
      241 aagaaaaaga aaaagcaatc acagcttatt aacgattcgt gtttaaataa tcaattgaaa
      301 ggaaaataga agatttttga aaaaaagtta ttggtattta ttaacttaat atatcagtaa
      361 catcaagaga aagaaactga attataaaca gagtatgtga agcaattgca attgcaatta
      421 ttagagatcc agaaaactga attgtgcttg aataccactt gtttaccggc gaacgtggcg
      481 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
      541 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
      601 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      661 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      721 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
      781 aattgggtac cgaattcgag ctcgtttggc ggccgctcta gaactagtgg atccagatct
      841 tgatccataa cttcgtatag catacattat acgaagttat agatccagta ctaataggaa
      901 ttgatttgga tggtataaac ggaaacaaaa aaaagagctg gtactacttt ctttaaaatt
      961 attttattat ttgattttat ttaatagtat atattatatt ttgaacgtag attattttgt
     1021 tgaaagttgc tgtagtgcca ttgattcgta acactaattc tgtattagtc attcctcttg
     1081 tttgatagta tccaaaaaaa cggctatttt tttgcaatct tatttcctgc atattataca
     1141 gataacataa tgaaagaaaa aatctttttt tttgttcttc aatgatgatt tcaaccattc
     1201 ttttaaacat tgatcaattc ctgagcaaca accccataca cactggttta tatatcgccc
     1261 cttttacagt tgaagaaaga aatagaaata gaaatagcaa acaaaagata tgacagtcaa
     1321 cactaagacc tatagtgaga gagcagaaac tcatgcctca ccagtagcac aacgattatt
     1381 tcgattaatg gaactgaaga aaaccaattt atgtgcatca attgatgttg ataccactaa
     1441 ggaattcctt gaattaattg ataaattggg tccttatgta tgcttaatca agactcatat
     1501 tgatataatc aatgattttt cctatgaatc cactattgaa ccattattag aactttcacg
     1561 taaacatcaa tttatgattt ttgaagatag aaaatttgct gatattggta ataccgtgaa
     1621 gaaacaatat attggtggag tttataaaat tagtagttgg gcagatatta ctaatgctca
     1681 tggtgtcact gggaatggag tagttgaagg attaaaacag ggagctaaag aaaccaccac
     1741 caaccaagag ccaagagggt tattgatgtt agctgaatta tcatcagtgg gatcattagc
     1801 atatggagaa tattctcaaa aaactgttga aattgctaaa tccgataagg aatttgttat
     1861 tggatttatt gcccaacgtg atatgggtgg acaagaagaa ggatttgatt ggcttattat
     1921 gacacctgga gttggattag atgataaagg tgatggatta ggacaacaat atagaactgt
     1981 tgatgaagtt gttagcactg gaactgatat tatcattgtt ggtagaggat tgtttggtaa
     2041 aggaagagat ccagatattg aaggtaaaag gtatagagat gctggttgga atgcttattt
     2101 gaaaaaaact ggccaattat aaatgtgaag ggggagattt tcactttatt agatttgtat
     2161 atatgtataa taaataaata aataagttaa ataaataatt aaataagggt ggtaattatt
     2221 actatttaca atcaaaggtg gtccttctag aagatctgct agcaattgtc tattccaagc
     2281 ctgtggcttt ttttggtgat tcacaataca taaattggca aatacctctt ctatagactt
     2341 taacctttga tgaaatggta tggataaacc acttttgtgg tcagggtata tatattttta
     2401 ttccaatctg gcctttataa attcaaattt ccaaggggac tctgctaaac cttctgatat
     2461 aactataact ctaataatgg ctgtaaaaag tgtgaaaaaa agatgatatt cacaaaaaag
     2521 ataaagggga tttcatattg ggtatctaga tttaaacatc gtatttcaac aatgaaacta
     2581 gaatcaatac gaaatccagc gcacaactgt gttcattccc ttgggaatta tactgagtaa
     2641 tgggaagaat gtgcaaaaaa tacccgtgtc ttatataatt tccatttgtg tagggcttaa
     2701 aagtggacaa cttcaggtta cgaaaccacg aaaatgtcta aacgtttgat caaattcgta
     2761 actcatggaa aatcacagca acaaattaag gatggatata tccggtactt acaaataaaa
     2821 acactaaaga agtataaatt ttgaactgaa attctataga aaatcttttt ttttgggtaa
     2881 aattatgcga ttgtggctca tagtaacgta atcatacagt tcaacaaaca tcattttgtc
     2941 aaaggatttc taatcccatt ctttttctac ttggcttttg agtatatata tatacatcct
     3001 ttgttcatca tcatgttcga ttgaattttt ttcttggttg ttattagatt gtctacttga
     3061 gtttttcttg gtgcaaattg tggtgttttt ctaacaaatt tttcaacatt taaagtattt
     3121 ctggaacttt tctacaagga acgacataga tccattaatg cgcccctcta aaacaatacc
     3181 cataatgtct tcttaagtat gcatctgctt ataaatgttt ctggatttac tttagttaga
     3241 atgtcgacac gacaatgaac tattgctgtg gatcacgtgc gatcactgaa ttatgagttt
     3301 ctttttattt ttctttttgg gctttttttt gcaatttttt ttttccacct ttgctaattg
     3361 cgtagtcaca tataatgtat acaatttttg caactctgac actataattg atccaataat
     3421 aaagtatata atagatcatc aagtatacgt aatctcccca aaaaaaaatt gcataaaata
     3481 aaatctataa acctttttta gatttttagt tgttcctttt ttctttctat tcttcaactt
     3541 ccaagtgttg tcactttctc cctgttaact atatttaaaa gtaaataccc tcccccccgg
     3601 gtatactcat taatcaataa ttatgtctaa cttgttgacc gttcaccaaa acttgccagc
     3661 tttgccagtt gatgctacct ctgatgaagt tagaaaaaac ttgatggata tgtttagaga
     3721 tagacaagcc ttctctgaac acacctggaa aatgttgttg tctgtttgta gatcctgggc
     3781 tgcttggtgt aaattgaaca acagaaaatg gttcccagcc gaaccagaag atgttagaga
     3841 ttacttgttg tacttgcaag ctagaggttt ggctgttaaa accattcaac aacacttggg
     3901 tcaattgaac atgttgcaca gaagatccgg tttgccaaga ccatctgatt ctaacgctgt
     3961 ttctttggtt atgagaagaa ttagaaaaga aaacgttgat gctggtgaaa gagctaaaca
     4021 agccttgtag gtatagacac tgaagaaaaa aaaattctat cattgttatg ttgtgatctt
     4081 tgatctttag ttgtcgggtt aacacctgcc aattggatca atacatcaat caattaattc
     4141 taatcttgaa aaaaaaattc attttactaa caattttttc ttttatttag ggctttcgaa
     4201 agaaccgatt tcgatcaagt tagatccttg atggaaaact ctgatagatg tcaagatatt
     4261 agaaacttgg ctttcttggg tattgcttac aacaccttgt tgagaattgc tgaaattgct
     4321 agaattagag ttaaagatat ttccagaacc gatggtggta gaatgttgat tcacattggt
     4381 agaaccaaaa ccttggtttc taccgctggt gttgaaaaag ccttgtcttt gggtgttacc
     4441 aaattggttg aaagatggat ttctgtttct ggtgttgctg atgatccaaa caactacttg
     4501 ttctgtagag ttagaaaaaa cggtgttgct gctccatctg ctacctctca attatctacc
     4561 agagctttgg aaggtatttt cgaagctacc cacagattga tttacggtgc taaagatgat
     4621 tctggtcaaa gatacttggc ttggtctggt cactctgcta gagttggtgc tgctagagat
     4681 atggctagag ccggcgtttc tattccagaa attatgcaag ctggtggttg gaccaacgtt
     4741 aacattgtta tgaactacat tagaaacttg gattctgaaa ccggtgctat ggttagattg
     4801 ttggaagatg gtgattaatt tatacctagg taagcaaata gctaaattat atacgaatta
     4861 atattatgat taagtgttta cgtgagtgcg atatttttat tactatctta tacagttgta
     4921 tatactctat aaaatgagtt gtctattaat taacgcgatg agtccctatt tattttttta
     4981 tagttatgtt agtattaaga acgttattta tatttcaaat taattatcca tggataactt
     5041 cgtatagcat acattatacg aagttattag ggataacagg gtaatctgca ggaagctcta
     5101 gaactagtgg atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc
     5161 tcgagggggg gccggaagga cgatgaagga gagaaagtag taagacaata tgactttgga
     5221 cagacatgag ataaaagttt taaaggacaa gaaaaggttc gggttaaaag atgagtattg
     5281 tattttttca aaatatcaat gctttttctg ttcttgggtt agtggtacca ccattggtga
     5341 taaaaccttt ggggagcctc ctccgagctt tctctttgtt actttttttc tcaactgata
     5401 cttgtcattc ttactacaat tacaggccta gccggcccaa cataggagcc ggaagcataa
     5461 agtgtaaagc ctggggtgcc taatgagtga ggtaactcac attaattgcg ttgcgctcac
     5521 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5581 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctcggcccc cctgacgagc
     5821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5881 aggcgttccc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     6001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6601 gttcatccat agttgcctga ctgcccgtcg tgtagataac tacgatacgg gagggcttac
     6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6961 gaaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg
     7561 ctataaaaat aattataatt taaatttttt aatataaata tataaattaa aaatagaaag
     7621 taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg
     7681 gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg
     7741 aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt
     7801 acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata
     7861 tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc
     7921 cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa
     7981 ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag
     8041 ttacaggcaa gcgatccgtc ctaagaaacc attattatca tgacattaac ctataaaaat
     8101 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     8161 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     8221 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     8281 tcagagcaga ttgtactgag agtgcaccat aaacgacatt actatatata taatatagga
     8341 agcatttaat agacagcatc gtaatatatg tgtactttgc agttatgacg ccagatggca
     8401 gtagtggaag atattcttta ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg
     8461 gagcttttct ttttttgccg attaagaatt aattcggtcg aaaaaagaaa aggagagggc
     8521 caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag
     8581 gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
     8641 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
     8701 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
     8761 tcaagtcttg taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg
     8821 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
     8881 gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
     8941 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
     9001 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     9061 tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg
     9121 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     9181 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     9241 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     9301 ttgtgcactt gcctgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     9361 aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     9421 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     9481 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggac
//
