LOCUS       62056_8465        9771 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pDIS-NAT1-Clox, complete sequence.
ACCESSION   MK652134
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9771)
  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 9771)
  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 9771)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE 
            (2019) In press"
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 13
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9771
                     /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"
     terminator      complement(1291..1488)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1506..2069)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(2089..2432)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2496..3827
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3855..4258,4423..5050)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59800.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4259..4422
                     /note="modified TUB2 intron sequence"
     3'UTR           5063..5139
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5194..5253
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5266..5299)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5379..5395)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5406..5655)
                     /label=NEUT5L 5'
                     /note="NEUT5L 5'"
     promoter        complement(5670..5700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5715..5736)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6024..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6786..7643)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7644..7748)
                     /label=AmpR promoter
     misc_feature    7785..8288
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        8544..8824
                     /label=TRP1 promoter
     CDS             8825..9496
                     /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 ctgagctcat cccattcatt ccatcataaa atgtcgacac
     1261 tggatggcgg cgttagtatc gaatcgacag cagtatagcg accagcattc acatacgatt
     1321 gacgcatgat attactttct gcgcacttaa cttcgcatct gggcagatga tgtcgaggcg
     1381 aaaaaaaata taaatcacgc taacatttga ttaaaataga acaactacaa tataaaaaaa
     1441 ctatacaaat gacaagttct tgaaaacaag aatcttttta ttgtcagtac tgactcgagt
     1501 tattatggac atggcataga catatacaaa gcttgttcac catcggaagc agtaccatcg
     1561 tataaagcag tatccaaacc acacaaagtg aaacccattc ttctataagc atgaatagct
     1621 ggagcattaa cattggtaac ttccaaccac aaatgaccag cacctctttc tctggcgaat
     1681 tcagtagcca aacccatcaa agctctacca acaccatgac ctctatgttc tggagcaact
     1741 tcaatatctt caacagtcaa tcttctgttc caaccagaat aagaaacaac aacgaaacca
     1801 gccaaatcac catcatcacc ataagcaacg aaagttctag aatctggatc accatcttca
     1861 ccagcatcgg attcatcatc ggattcatca tctgggaaaa ccttagtcaa tggtggatca
     1921 actggaactt ctctcaaagt gaaaccatca ccagtagcag taactctaaa aacagtatcg
     1981 gtagtgaaag aaccatccaa agcttcaata gcttcagcat cacctggaac agaagttctg
     2041 tatctataag cagtatcatc caaagtagta gacataattg taggatccgg ttgtttatgt
     2101 tcggatgtga tgtgagaact gtatcctagc aagattttaa aaggaagtat atgaaagaag
     2161 aacctcagtg gcaaatccta accttttata tttctctaca ggggcgcggc gtggggacaa
     2221 ttcaacgcgt ctgtgagggg agcgtttccc tgctcgcagg tctgcagcga ggagccgtaa
     2281 tttttgcttc gcgccgtgcg gccatcaaaa tgtatggatg caaatgatta tacatgggga
     2341 tgtatgggct aaatgtacgg gcgacagtca catcatgccc ctgagctgcg cacgtcaaga
     2401 ctgtcaagga gggtattctg ggcctccatg tcgctggccg ggtgacccgg cggggacgag
     2461 gcaagcttga tggaagttcc tatactttcg ctagcaattg tctattccaa gcctgtggct
     2521 ttttttggtg attcacaata cataaattgg caaatacctc ttctatagac tttaaccttt
     2581 gatgaaatgg tatggataaa ccacttttgt ggtcagggta tatatatttt tattccaatc
     2641 tggcctttat aaattcaaat ttccaagggg actctgctaa accttctgat ataactataa
     2701 ctctaataat ggctgtaaaa agtgtgaaaa aaagatgata ttcacaaaaa agataaaggg
     2761 gatttcatat tgggtatcta gatttaaaca tcgtatttca acaatgaaac tagaatcaat
     2821 acgaaatcca gcgcacaact gtgttcattc ccttgggaat tatactgagt aatgggaaga
     2881 atgtgcaaaa aatacccgtg tcttatataa tttccatttg tgtagggctt aaaagtggac
     2941 aacttcaggt tacgaaacca cgaaaatgtc taaacgtttg atcaaattcg taactcatgg
     3001 aaaatcacag caacaaatta aggatggata tatccggtac ttacaaataa aaacactaaa
     3061 gaagtataaa ttttgaactg aaattctata gaaaatcttt ttttttgggt aaaattatgc
     3121 gattgtggct catagtaacg taatcataca gttcaacaaa catcattttg tcaaaggatt
     3181 tctaatccca ttctttttct acttggcttt tgagtatata tatatacatc ctttgttcat
     3241 catcatgttc gattgaattt ttttcttggt tgttattaga ttgtctactt gagtttttct
     3301 tggtgcaaat tgtggtgttt ttctaacaaa tttttcaaca tttaaagtat ttctggaact
     3361 tttctacaag gaacgacata gatccattaa tgcgcccctc taaaacaata cccataatgt
     3421 cttcttaagt atgcatctgc ttataaatgt ttctggattt actttagtta gaatgtcgac
     3481 acgacaatga actattgctg tggatcacgt gcgatcactg aattatgagt ttctttttat
     3541 ttttcttttt gggctttttt ttgcaatttt ttttttccac ctttgctaat tgcgtagtca
     3601 catataatgt atacaatttt tgcaactctg acactataat tgatccaata ataaagtata
     3661 taatagatca tcaagtatac gtaatctccc caaaaaaaaa ttgcataaaa taaaatctat
     3721 aaaccttttt tagattttta gttgttcctt ttttctttct attcttcaac ttccaagtgt
     3781 tgtcactttc tccctgttaa ctatatttaa aagtaaatac cctccccccc gggtatactc
     3841 attaatcaat aattatgtct aacttgttga ccgttcacca aaacttgcca gctttgccag
     3901 ttgatgctac ctctgatgaa gttagaaaaa acttgatgga tatgtttaga gatagacaag
     3961 ccttctctga acacacctgg aaaatgttgt tgtctgtttg tagatcctgg gctgcttggt
     4021 gtaaattgaa caacagaaaa tggttcccag ccgaaccaga agatgttaga gattacttgt
     4081 tgtacttgca agctagaggt ttggctgtta aaaccattca acaacacttg ggtcaattga
     4141 acatgttgca cagaagatcc ggtttgccaa gaccatctga ttctaacgct gtttctttgg
     4201 ttatgagaag aattagaaaa gaaaacgttg atgctggtga aagagctaaa caagccttgt
     4261 aggtatagac actgaagaaa aaaaaattct atcattgtta tgttgtgatc tttgatcttt
     4321 agttgtcggg ttaacacctg ccaattggat caatacatca atcaattaat tctaatcttg
     4381 aaaaaaaaat tcattttact aacaattttt tcttttattt agggctttcg aaagaaccga
     4441 tttcgatcaa gttagatcct tgatggaaaa ctctgataga tgtcaagata ttagaaactt
     4501 ggctttcttg ggtattgctt acaacacctt gttgagaatt gctgaaattg ctagaattag
     4561 agttaaagat atttccagaa ccgatggtgg tagaatgttg attcacattg gtagaaccaa
     4621 aaccttggtt tctaccgctg gtgttgaaaa agccttgtct ttgggtgtta ccaaattggt
     4681 tgaaagatgg atttctgttt ctggtgttgc tgatgatcca aacaactact tgttctgtag
     4741 agttagaaaa aacggtgttg ctgctccatc tgctacctct caattatcta ccagagcttt
     4801 ggaaggtatt ttcgaagcta cccacagatt gatttacggt gctaaagatg attctggtca
     4861 aagatacttg gcttggtctg gtcactctgc tagagttggt gctgctagag atatggctag
     4921 agccggcgtt tctattccag aaattatgca agctggtggt tggaccaacg ttaacattgt
     4981 tatgaactac attagaaact tggattctga aaccggtgct atggttagat tgttggaaga
     5041 tggtgattaa tttataccta ggtaagcaaa tagctaaatt atatacgaat taatattatg
     5101 attaagtgtt tacgtgagtg cgatattttt attactatct tatacagttg tatatactct
     5161 ataaaatgag ttgtctatta attaacgcga tgagtcccta tttatttttt tatagttatg
     5221 ttagtattaa gaacgttatt tatatttcaa attaattatc catggataac ttcgtatagc
     5281 atacattata cgaagttatt agggataaca gggtaatctg caggaagctc tagaactagt
     5341 ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga cctcgagggg
     5401 gggccggaag gacgatgaag gagagaaagt agtaagacaa tatgactttg gacagacatg
     5461 agataaaagt tttaaaggac aagaaaaggt tcgggttaaa agatgagtat tgtatttttt
     5521 caaaatatca atgctttttc tgttcttggg ttagtggtac caccattggt gataaaacct
     5581 ttggggagcc tcctccgagc tttctctttg ttactttttt tctcaactga tacttgtcat
     5641 tcttactaca attacaggcc tagccggccc aacataggag ccggaagcat aaagtgtaaa
     5701 gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct
     5761 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5821 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5881 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6001 aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga gcatcacaaa
     6061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttc
     6121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6181 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6421 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6841 atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt accatctggc
     6901 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgaaaaaaa
     7201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7741 gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa
     7801 ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa
     7861 gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc
     7921 caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt
     7981 catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg
     8041 ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt
     8101 acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc
     8161 ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac
     8221 agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc
     8281 aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8341 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8401 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     8461 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     8521 gattgtactg agagtgcacc ataaacgaca ttactatata tataatatag gaagcattta
     8581 atagacagca tcgtaatata tgtgtacttt gcagttatga cgccagatgg cagtagtgga
     8641 agatattctt tattgaaaaa tagcttgtca ccttacgtac aatcttgatc cggagctttt
     8701 ctttttttgc cgattaagaa ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg
     8761 agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg
     8821 gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt
     8881 gcagagcaca gaggccgcag aatgtgctct agattccgat gctgacttgc tgggtattat
     8941 atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct
     9001 tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa
     9061 tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact
     9121 gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag
     9181 actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc
     9241 gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc
     9301 gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg
     9361 tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga
     9421 tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt
     9481 caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac
     9541 ttgcctgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     9601 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     9661 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     9721 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga c
//
