LOCUS       V016521                 9185 bp    DNA     circular SYN 17-JUL-2019
DEFINITION  Exported.
ACCESSION   V016521
VERSION     V016521
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9185)
  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 9185)
  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 9185)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 12
            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..9185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             62..772
                     /label="GFP"
                     /note="green fluorescent protein"
     regulatory      2210..2465
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     2475..2491
                     /label="SK primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2505..2538
                     /product="loxP"
     CDS             2968..3777
                     /gene="URA3"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Candida
                     albicans (strain SC5314 / ATCC MYA-2876). Accession#:
                     P13649"
     regulatory      3922..5253
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(5281..5684,5849..6476)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label="Cre"
                     /note="Cre recombinase"
                     /protein_id="QDK59728.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          5685..5848
                     /note="modified TUB2 intron sequence"
     3'UTR           6489..6619
                     /label="Saccharomyces cerevisiae ADH1 3'UTR region"
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      6620..6679
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(6692..6725)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(6823..6841)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(7099..7687)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7861..8718)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8719..8823)
                     /label="AmpR promoter"
     promoter        9169..9185
                     /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 tagccgacca
      601 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
      661 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
      721 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aaagatcctc
      781 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
      841 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
      901 gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
      961 tactttatgt tatggtgttc aatgtttttc aagataccca gatcatatga aacaacatga
     1021 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
     1081 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
     1141 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
     1201 atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa
     1261 agctaacttc aaaactagac acaacattga agatggttct gttcaattag ctgaccatta
     1321 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
     1381 cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
     1441 atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaaa gatcctctaa
     1501 aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa
     1561 tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac
     1621 cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac
     1681 tttatgttat ggtgttcaat gtttttcaag atacccagat catatgaaac aacatgactt
     1741 tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga
     1801 cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat
     1861 cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata
     1921 caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagc
     1981 taacttcaaa actagacaca acattgaaga tggttctgtt caattagctg accattatca
     2041 acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac
     2101 tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt
     2161 tgttactgct gctggtatta cccatggtat ggatgaattg tacaaataag gcgcgccatt
     2221 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     2281 attacccggg aatctcggtc gtaatgattt ctataatgac gaaaaaaaaa aaattggaaa
     2341 gaaaaagctt catggccttt ataaaaagga actatccaat acctcgccag aaccaagtaa
     2401 cagtatttta cggggcacaa atcaagaaca ataagacagg actgtaaaga tggacgcata
     2461 gatccggcgg ccgctctaga actagtggat ccagatcttg atccataact tcgtatagca
     2521 tacattatac gaagttatag atccagtact aataggaatt gatttggatg gtataaacgg
     2581 aaacaaaaaa aagagctggt actactttct ttaaaattat tttattattt gattttattt
     2641 aatagtatat attatatttt gaacgtagat tattttgttg aaagttgctg tagtgccatt
     2701 gattcgtaac actaattctg tattagtcat tcctcttgtt tgatagtatc caaaaaaacg
     2761 gctatttttt tgcaatctta tttcctgcat attatacaga taacataatg aaagaaaaaa
     2821 tctttttttt tgttcttcaa tgatgatttc aaccattctt ttaaacattg atcaattcct
     2881 gagcaacaac cccatacaca ctggtttata tatcgcccct tttacagttg aagaaagaaa
     2941 tagaaataga aatagcaaac aaaagatatg acagtcaaca ctaagaccta tagtgagaga
     3001 gcagaaactc atgcctcacc agtagcacaa cgattatttc gattaatgga actgaagaaa
     3061 accaatttat gtgcatcaat tgatgttgat accactaagg aattccttga attaattgat
     3121 aaattgggtc cttatgtatg cttaatcaag actcatattg atataatcaa tgatttttcc
     3181 tatgaatcca ctattgaacc attattagaa ctttcacgta aacatcaatt tatgattttt
     3241 gaagatagaa aatttgctga tattggtaat accgtgaaga aacaatatat tggtggagtt
     3301 tataaaatta gtagttgggc agatattact aatgctcatg gtgtcactgg gaatggagta
     3361 gttgaaggat taaaacaggg agctaaagaa accaccacca accaagagcc aagagggtta
     3421 ttgatgttag ctgaattatc atcagtggga tcattagcat atggagaata ttctcaaaaa
     3481 actgttgaaa ttgctaaatc cgataaggaa tttgttattg gatttattgc ccaacgtgat
     3541 atgggtggac aagaagaagg atttgattgg cttattatga cacctggagt tggattagat
     3601 gataaaggtg atggattagg acaacaatat agaactgttg atgaagttgt tagcactgga
     3661 actgatatta tcattgttgg tagaggattg tttggtaaag gaagagatcc agatattgaa
     3721 ggtaaaaggt atagagatgc tggttggaat gcttatttga aaaaaactgg ccaattataa
     3781 atgtgaaggg ggagattttc actttattag atttgtatat atgtataata aataaataaa
     3841 taagttaaat aaataattaa ataagggtgg taattattac tatttacaat caaaggtggt
     3901 ccttctagaa gatctgctag caattgtcta ttccaagcct gtggcttttt ttggtgattc
     3961 acaatacata aattggcaaa tacctcttct atagacttta acctttgatg aaatggtatg
     4021 gataaaccac ttttgtggtc agggtatata tatttttatt ccaatctggc ctttataaat
     4081 tcaaatttcc aaggggactc tgctaaacct tctgatataa ctataactct aataatggct
     4141 gtaaaaagtg tgaaaaaaag atgatattca caaaaaagat aaaggggatt tcatattggg
     4201 tatctagatt taaacatcgt atttcaacaa tgaaactaga atcaatacga aatccagcgc
     4261 acaactgtgt tcattccctt gggaattata ctgagtaatg ggaagaatgt gcaaaaaata
     4321 cccgtgtctt atataatttc catttgtgta gggcttaaaa gtggacaact tcaggttacg
     4381 aaaccacgaa aatgtctaaa cgtttgatca aattcgtaac tcatggaaaa tcacagcaac
     4441 aaattaagga tggatatatc cggtacttac aaataaaaac actaaagaag tataaatttt
     4501 gaactgaaat tctatagaaa atcttttttt ttgggtaaaa ttatgcgatt gtggctcata
     4561 gtaacgtaat catacagttc aacaaacatc attttgtcaa aggatttcta atcccattct
     4621 ttttctactt ggcttttgag tatatatata tacatccttt gttcatcatc atgttcgatt
     4681 gaattttttt cttggttgtt attagattgt ctacttgagt ttttcttggt gcaaattgtg
     4741 gtgtttttct aacaaatttt tcaacattta aagtatttct ggaacttttc tacaaggaac
     4801 gacatagatc cattaatgcg cccctctaaa acaataccca taatgtcttc ttaagtatgc
     4861 atctgcttat aaatgtttct ggatttactt tagttagaat gtcgacacga caatgaacta
     4921 ttgctgtgga tcacgtgcga tcactgaatt atgagtttct ttttattttt ctttttgggc
     4981 ttttttttgc aatttttttt ttccaccttt gctaattgcg tagtcacata taatgtatac
     5041 aatttttgca actctgacac tataattgat ccaataataa agtatataat agatcatcaa
     5101 gtatacgtaa tctccccaaa aaaaaattgc ataaaataaa atctataaac cttttttaga
     5161 tttttagttg ttcctttttt ctttctattc ttcaacttcc aagtgttgtc actttctccc
     5221 tgttaactat atttaaaagt aaataccctc ccccccgggt atactcatta atcaataatt
     5281 atgtctaact tgttgaccgt tcaccaaaac ttgccagctt tgccagttga tgctacctct
     5341 gatgaagtta gaaaaaactt gatggatatg tttagagata gacaagcctt ctctgaacac
     5401 acctggaaaa tgttgttgtc tgtttgtaga tcctgggctg cttggtgtaa attgaacaac
     5461 agaaaatggt tcccagccga accagaagat gttagagatt acttgttgta cttgcaagct
     5521 agaggtttgg ctgttaaaac cattcaacaa cacttgggtc aattgaacat gttgcacaga
     5581 agatccggtt tgccaagacc atctgattct aacgctgttt ctttggttat gagaagaatt
     5641 agaaaagaaa acgttgatgc tggtgaaaga gctaaacaag ccttgtaggt atagacactg
     5701 aagaaaaaaa aattctatca ttgttatgtt gtgatctttg atctttagtt gtcgggttaa
     5761 cacctgccaa ttggatcaat acatcaatca attaattcta atcttgaaaa aaaaattcat
     5821 tttactaaca attttttctt ttatttaggg ctttcgaaag aaccgatttc gatcaagtta
     5881 gatccttgat ggaaaactct gatagatgtc aagatattag aaacttggct ttcttgggta
     5941 ttgcttacaa caccttgttg agaattgctg aaattgctag aattagagtt aaagatattt
     6001 ccagaaccga tggtggtaga atgttgattc acattggtag aaccaaaacc ttggtttcta
     6061 ccgctggtgt tgaaaaagcc ttgtctttgg gtgttaccaa attggttgaa agatggattt
     6121 ctgtttctgg tgttgctgat gatccaaaca actacttgtt ctgtagagtt agaaaaaacg
     6181 gtgttgctgc tccatctgct acctctcaat tatctaccag agctttggaa ggtattttcg
     6241 aagctaccca cagattgatt tacggtgcta aagatgattc tggtcaaaga tacttggctt
     6301 ggtctggtca ctctgctaga gttggtgctg ctagagatat ggctagagcc ggcgtttcta
     6361 ttccagaaat tatgcaagct ggtggttgga ccaacgttaa cattgttatg aactacatta
     6421 gaaacttgga ttctgaaacc ggtgctatgg ttagattgtt ggaagatggt gattaattta
     6481 tacctaggta agcaaatagc taaattatat acgaattaat attatgatta agtgtttacg
     6541 tgagtgcgat atttttatta ctatcttata cagttgtata tactctataa aatgagttgt
     6601 ctattaatta acgcgatgag tccctattta tttttttata gttatgttag tattaagaac
     6661 gttatttata tttcaaatta attatccatg gataacttcg tatagcatac attatacgaa
     6721 gttattaggg ataacagggt aatctgcagg aaaaacgagc tcgaattcat cgatgatatc
     6781 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     6841 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7261 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7501 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8821 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8881 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     8941 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     9001 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     9061 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
     9121 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     9181 ctata
//
