LOCUS       V016152                 7123 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V016152
VERSION     V016152
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7123)
  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 7123)
  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 7123)
  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..7123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..93
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             97..123
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             127..153
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     regulatory      157..412
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     misc_feature    426..459
                     /label="loxP"
                     /note="loxP"
     CDS             889..1698
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      1843..3174
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3202..3605,3770..4397)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59776.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3606..3769
                     /note="modified TUB2 intron sequence"
     3'UTR           4410..4540
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4541..4600
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(4613..4646)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4744..4762)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5782..6639)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6640..6744)
                     /label="AmpR promoter"
     promoter        7090..7108
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaagcttcgt acgctgcagg tcgacgggcg gccgctctag aactagtggg tcgacggatc
       61 cccgggtacc catacgatgt tcctgactat gcgggctatc cctatgacgt cccggactat
      121 gcaggatatc catatgacgt tccagattac gcttagggcg cgccattata agtaaatgca
      181 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat
      241 ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat
      301 ggcctttata aaaaggaact atccaatacc tcgccagaac caagtaacag tattttacgg
      361 ggcacaaatc aagaacaata agacaggact gtaaagatgg acgcatagat cccagatctt
      421 gatccataac ttcgtatagc atacattata cgaagttata gatccagtac taataggaat
      481 tgatttggat ggtataaacg gaaacaaaaa aaagagctgg tactactttc tttaaaatta
      541 ttttattatt tgattttatt taatagtata tattatattt tgaacgtaga ttattttgtt
      601 gaaagttgct gtagtgccat tgattcgtaa cactaattct gtattagtca ttcctcttgt
      661 ttgatagtat ccaaaaaaac ggctattttt ttgcaatctt atttcctgca tattatacag
      721 ataacataat gaaagaaaaa atcttttttt ttgttcttca atgatgattt caaccattct
      781 tttaaacatt gatcaattcc tgagcaacaa ccccatacac actggtttat atatcgcccc
      841 ttttacagtt gaagaaagaa atagaaatag aaatagcaaa caaaagatat gacagtcaac
      901 actaagacct atagtgagag agcagaaact catgcctcac cagtagcaca acgattattt
      961 cgattaatgg aactgaagaa aaccaattta tgtgcatcaa ttgatgttga taccactaag
     1021 gaattccttg aattaattga taaattgggt ccttatgtat gcttaatcaa gactcatatt
     1081 gatataatca atgatttttc ctatgaatcc actattgaac cattattaga actttcacgt
     1141 aaacatcaat ttatgatttt tgaagataga aaatttgctg atattggtaa taccgtgaag
     1201 aaacaatata ttggtggagt ttataaaatt agtagttggg cagatattac taatgctcat
     1261 ggtgtcactg ggaatggagt agttgaagga ttaaaacagg gagctaaaga aaccaccacc
     1321 aaccaagagc caagagggtt attgatgtta gctgaattat catcagtggg atcattagca
     1381 tatggagaat attctcaaaa aactgttgaa attgctaaat ccgataagga atttgttatt
     1441 ggatttattg cccaacgtga tatgggtgga caagaagaag gatttgattg gcttattatg
     1501 acacctggag ttggattaga tgataaaggt gatggattag gacaacaata tagaactgtt
     1561 gatgaagttg ttagcactgg aactgatatt atcattgttg gtagaggatt gtttggtaaa
     1621 ggaagagatc cagatattga aggtaaaagg tatagagatg ctggttggaa tgcttatttg
     1681 aaaaaaactg gccaattata aatgtgaagg gggagatttt cactttatta gatttgtata
     1741 tatgtataat aaataaataa ataagttaaa taaataatta aataagggtg gtaattatta
     1801 ctatttacaa tcaaaggtgg tccttctaga agatctgcta gcaattgtct attccaagcc
     1861 tgtggctttt tttggtgatt cacaatacat aaattggcaa atacctcttc tatagacttt
     1921 aacctttgat gaaatggtat ggataaacca cttttgtggt cagggtatat atatttttat
     1981 tccaatctgg cctttataaa ttcaaatttc caaggggact ctgctaaacc ttctgatata
     2041 actataactc taataatggc tgtaaaaagt gtgaaaaaaa gatgatattc acaaaaaaga
     2101 taaaggggat ttcatattgg gtatctagat ttaaacatcg tatttcaaca atgaaactag
     2161 aatcaatacg aaatccagcg cacaactgtg ttcattccct tgggaattat actgagtaat
     2221 gggaagaatg tgcaaaaaat acccgtgtct tatataattt ccatttgtgt agggcttaaa
     2281 agtggacaac ttcaggttac gaaaccacga aaatgtctaa acgtttgatc aaattcgtaa
     2341 ctcatggaaa atcacagcaa caaattaagg atggatatat ccggtactta caaataaaaa
     2401 cactaaagaa gtataaattt tgaactgaaa ttctatagaa aatctttttt tttgggtaaa
     2461 attatgcgat tgtggctcat agtaacgtaa tcatacagtt caacaaacat cattttgtca
     2521 aaggatttct aatcccattc tttttctact tggcttttga gtatatatat atacatcctt
     2581 tgttcatcat catgttcgat tgaatttttt tcttggttgt tattagattg tctacttgag
     2641 tttttcttgg tgcaaattgt ggtgtttttc taacaaattt ttcaacattt aaagtatttc
     2701 tggaactttt ctacaaggaa cgacatagat ccattaatgc gcccctctaa aacaataccc
     2761 ataatgtctt cttaagtatg catctgctta taaatgtttc tggatttact ttagttagaa
     2821 tgtcgacacg acaatgaact attgctgtgg atcacgtgcg atcactgaat tatgagtttc
     2881 tttttatttt tctttttggg cttttttttg caattttttt tttccacctt tgctaattgc
     2941 gtagtcacat ataatgtata caatttttgc aactctgaca ctataattga tccaataata
     3001 aagtatataa tagatcatca agtatacgta atctccccaa aaaaaaattg cataaaataa
     3061 aatctataaa ccttttttag atttttagtt gttccttttt tctttctatt cttcaacttc
     3121 caagtgttgt cactttctcc ctgttaacta tatttaaaag taaataccct cccccccggg
     3181 tatactcatt aatcaataat tatgtctaac ttgttgaccg ttcaccaaaa cttgccagct
     3241 ttgccagttg atgctacctc tgatgaagtt agaaaaaact tgatggatat gtttagagat
     3301 agacaagcct tctctgaaca cacctggaaa atgttgttgt ctgtttgtag atcctgggct
     3361 gcttggtgta aattgaacaa cagaaaatgg ttcccagccg aaccagaaga tgttagagat
     3421 tacttgttgt acttgcaagc tagaggtttg gctgttaaaa ccattcaaca acacttgggt
     3481 caattgaaca tgttgcacag aagatccggt ttgccaagac catctgattc taacgctgtt
     3541 tctttggtta tgagaagaat tagaaaagaa aacgttgatg ctggtgaaag agctaaacaa
     3601 gccttgtagg tatagacact gaagaaaaaa aaattctatc attgttatgt tgtgatcttt
     3661 gatctttagt tgtcgggtta acacctgcca attggatcaa tacatcaatc aattaattct
     3721 aatcttgaaa aaaaaattca ttttactaac aattttttct tttatttagg gctttcgaaa
     3781 gaaccgattt cgatcaagtt agatccttga tggaaaactc tgatagatgt caagatatta
     3841 gaaacttggc tttcttgggt attgcttaca acaccttgtt gagaattgct gaaattgcta
     3901 gaattagagt taaagatatt tccagaaccg atggtggtag aatgttgatt cacattggta
     3961 gaaccaaaac cttggtttct accgctggtg ttgaaaaagc cttgtctttg ggtgttacca
     4021 aattggttga aagatggatt tctgtttctg gtgttgctga tgatccaaac aactacttgt
     4081 tctgtagagt tagaaaaaac ggtgttgctg ctccatctgc tacctctcaa ttatctacca
     4141 gagctttgga aggtattttc gaagctaccc acagattgat ttacggtgct aaagatgatt
     4201 ctggtcaaag atacttggct tggtctggtc actctgctag agttggtgct gctagagata
     4261 tggctagagc cggcgtttct attccagaaa ttatgcaagc tggtggttgg accaacgtta
     4321 acattgttat gaactacatt agaaacttgg attctgaaac cggtgctatg gttagattgt
     4381 tggaagatgg tgattaattt atacctaggt aagcaaatag ctaaattata tacgaattaa
     4441 tattatgatt aagtgtttac gtgagtgcga tatttttatt actatcttat acagttgtat
     4501 atactctata aaatgagttg tctattaatt aacgcgatga gtccctattt atttttttat
     4561 agttatgtta gtattaagaa cgttatttat atttcaaatt aattatccat ggataacttc
     4621 gtatagcata cattatacga agttattagg gataacaggg taatctgcag gaaaaacgag
     4681 ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt
     4741 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     4801 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5761 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5821 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5881 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5941 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6001 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6061 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6121 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6181 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6241 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6301 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6361 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6421 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6481 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6541 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6601 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6661 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6721 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6781 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6841 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6901 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6961 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7021 ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
     7081 acacatacga tttaggtgac actatagaac gcggccgcca gct
//
