LOCUS       V016537                 7751 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V016537
VERSION     V016537
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7751)
  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 7751)
  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 7751)
  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..7751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             62..772
                     /label="GFP"
                     /note="green fluorescent protein"
     regulatory      776..1031
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     1041..1057
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1071..1104
                     /label="loxP"
                     /note="loxP"
     CDS             1534..2343
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      2488..3819
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3847..4250,4415..5042)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59740.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4251..4414
                     /note="modified TUB2 intron sequence"
     3'UTR           5055..5185
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5186..5245
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5258..5291)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5389..5407)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5665..6253)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6427..7284)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7285..7389)
                     /label="AmpR promoter"
     promoter        7735..7751
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggtgctg gcgcaggtgc
       61 ttctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
      121 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
      181 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
      241 cactacttta tgttatggtg ttcaatgttt ttcaagatac ccagatcata tgaaacaaca
      301 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
      361 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
      421 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
      481 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
      541 caaagctaac ttcaaaacta gacacaacat tgaagatggt tctgttcaat tagctgacca
      601 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
      661 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
      721 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aatagggcgc
      781 gccattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
      841 tcagttatta cccgggaatc tcggtcgtaa tgatttctat aatgacgaaa aaaaaaaaat
      901 tggaaagaaa aagcttcatg gcctttataa aaaggaacta tccaatacct cgccagaacc
      961 aagtaacagt attttacggg gcacaaatca agaacaataa gacaggactg taaagatgga
     1021 cgcatagatc cggcggccgc tctagaacta gtggatccag atcttgatcc ataacttcgt
     1081 atagcataca ttatacgaag ttatagatcc agtactaata ggaattgatt tggatggtat
     1141 aaacggaaac aaaaaaaaga gctggtacta ctttctttaa aattatttta ttatttgatt
     1201 ttatttaata gtatatatta tattttgaac gtagattatt ttgttgaaag ttgctgtagt
     1261 gccattgatt cgtaacacta attctgtatt agtcattcct cttgtttgat agtatccaaa
     1321 aaaacggcta tttttttgca atcttatttc ctgcatatta tacagataac ataatgaaag
     1381 aaaaaatctt tttttttgtt cttcaatgat gatttcaacc attcttttaa acattgatca
     1441 attcctgagc aacaacccca tacacactgg tttatatatc gcccctttta cagttgaaga
     1501 aagaaataga aatagaaata gcaaacaaaa gatatgacag tcaacactaa gacctatagt
     1561 gagagagcag aaactcatgc ctcaccagta gcacaacgat tatttcgatt aatggaactg
     1621 aagaaaacca atttatgtgc atcaattgat gttgatacca ctaaggaatt ccttgaatta
     1681 attgataaat tgggtcctta tgtatgctta atcaagactc atattgatat aatcaatgat
     1741 ttttcctatg aatccactat tgaaccatta ttagaacttt cacgtaaaca tcaatttatg
     1801 atttttgaag atagaaaatt tgctgatatt ggtaataccg tgaagaaaca atatattggt
     1861 ggagtttata aaattagtag ttgggcagat attactaatg ctcatggtgt cactgggaat
     1921 ggagtagttg aaggattaaa acagggagct aaagaaacca ccaccaacca agagccaaga
     1981 gggttattga tgttagctga attatcatca gtgggatcat tagcatatgg agaatattct
     2041 caaaaaactg ttgaaattgc taaatccgat aaggaatttg ttattggatt tattgcccaa
     2101 cgtgatatgg gtggacaaga agaaggattt gattggctta ttatgacacc tggagttgga
     2161 ttagatgata aaggtgatgg attaggacaa caatatagaa ctgttgatga agttgttagc
     2221 actggaactg atattatcat tgttggtaga ggattgtttg gtaaaggaag agatccagat
     2281 attgaaggta aaaggtatag agatgctggt tggaatgctt atttgaaaaa aactggccaa
     2341 ttataaatgt gaagggggag attttcactt tattagattt gtatatatgt ataataaata
     2401 aataaataag ttaaataaat aattaaataa gggtggtaat tattactatt tacaatcaaa
     2461 ggtggtcctt ctagaagatc tgctagcaat tgtctattcc aagcctgtgg ctttttttgg
     2521 tgattcacaa tacataaatt ggcaaatacc tcttctatag actttaacct ttgatgaaat
     2581 ggtatggata aaccactttt gtggtcaggg tatatatatt tttattccaa tctggccttt
     2641 ataaattcaa atttccaagg ggactctgct aaaccttctg atataactat aactctaata
     2701 atggctgtaa aaagtgtgaa aaaaagatga tattcacaaa aaagataaag gggatttcat
     2761 attgggtatc tagatttaaa catcgtattt caacaatgaa actagaatca atacgaaatc
     2821 cagcgcacaa ctgtgttcat tcccttggga attatactga gtaatgggaa gaatgtgcaa
     2881 aaaatacccg tgtcttatat aatttccatt tgtgtagggc ttaaaagtgg acaacttcag
     2941 gttacgaaac cacgaaaatg tctaaacgtt tgatcaaatt cgtaactcat ggaaaatcac
     3001 agcaacaaat taaggatgga tatatccggt acttacaaat aaaaacacta aagaagtata
     3061 aattttgaac tgaaattcta tagaaaatct ttttttttgg gtaaaattat gcgattgtgg
     3121 ctcatagtaa cgtaatcata cagttcaaca aacatcattt tgtcaaagga tttctaatcc
     3181 cattcttttt ctacttggct tttgagtata tatatataca tcctttgttc atcatcatgt
     3241 tcgattgaat ttttttcttg gttgttatta gattgtctac ttgagttttt cttggtgcaa
     3301 attgtggtgt ttttctaaca aatttttcaa catttaaagt atttctggaa cttttctaca
     3361 aggaacgaca tagatccatt aatgcgcccc tctaaaacaa tacccataat gtcttcttaa
     3421 gtatgcatct gcttataaat gtttctggat ttactttagt tagaatgtcg acacgacaat
     3481 gaactattgc tgtggatcac gtgcgatcac tgaattatga gtttcttttt atttttcttt
     3541 ttgggctttt ttttgcaatt ttttttttcc acctttgcta attgcgtagt cacatataat
     3601 gtatacaatt tttgcaactc tgacactata attgatccaa taataaagta tataatagat
     3661 catcaagtat acgtaatctc cccaaaaaaa aattgcataa aataaaatct ataaaccttt
     3721 tttagatttt tagttgttcc ttttttcttt ctattcttca acttccaagt gttgtcactt
     3781 tctccctgtt aactatattt aaaagtaaat accctccccc ccgggtatac tcattaatca
     3841 ataattatgt ctaacttgtt gaccgttcac caaaacttgc cagctttgcc agttgatgct
     3901 acctctgatg aagttagaaa aaacttgatg gatatgttta gagatagaca agccttctct
     3961 gaacacacct ggaaaatgtt gttgtctgtt tgtagatcct gggctgcttg gtgtaaattg
     4021 aacaacagaa aatggttccc agccgaacca gaagatgtta gagattactt gttgtacttg
     4081 caagctagag gtttggctgt taaaaccatt caacaacact tgggtcaatt gaacatgttg
     4141 cacagaagat ccggtttgcc aagaccatct gattctaacg ctgtttcttt ggttatgaga
     4201 agaattagaa aagaaaacgt tgatgctggt gaaagagcta aacaagcctt gtaggtatag
     4261 acactgaaga aaaaaaaatt ctatcattgt tatgttgtga tctttgatct ttagttgtcg
     4321 ggttaacacc tgccaattgg atcaatacat caatcaatta attctaatct tgaaaaaaaa
     4381 attcatttta ctaacaattt tttcttttat ttagggcttt cgaaagaacc gatttcgatc
     4441 aagttagatc cttgatggaa aactctgata gatgtcaaga tattagaaac ttggctttct
     4501 tgggtattgc ttacaacacc ttgttgagaa ttgctgaaat tgctagaatt agagttaaag
     4561 atatttccag aaccgatggt ggtagaatgt tgattcacat tggtagaacc aaaaccttgg
     4621 tttctaccgc tggtgttgaa aaagccttgt ctttgggtgt taccaaattg gttgaaagat
     4681 ggatttctgt ttctggtgtt gctgatgatc caaacaacta cttgttctgt agagttagaa
     4741 aaaacggtgt tgctgctcca tctgctacct ctcaattatc taccagagct ttggaaggta
     4801 ttttcgaagc tacccacaga ttgatttacg gtgctaaaga tgattctggt caaagatact
     4861 tggcttggtc tggtcactct gctagagttg gtgctgctag agatatggct agagccggcg
     4921 tttctattcc agaaattatg caagctggtg gttggaccaa cgttaacatt gttatgaact
     4981 acattagaaa cttggattct gaaaccggtg ctatggttag attgttggaa gatggtgatt
     5041 aatttatacc taggtaagca aatagctaaa ttatatacga attaatatta tgattaagtg
     5101 tttacgtgag tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg
     5161 agttgtctat taattaacgc gatgagtccc tatttatttt tttatagtta tgttagtatt
     5221 aagaacgtta tttatatttc aaattaatta tccatggata acttcgtata gcatacatta
     5281 tacgaagtta ttagggataa cagggtaatc tgcaggaaaa acgagctcga attcatcgat
     5341 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     5401 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     5461 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5521 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5581 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6301 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6361 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6421 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6481 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6601 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6841 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7081 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7261 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7381 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7441 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7501 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7561 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7621 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     7681 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     7741 gtgacactat a
//
