LOCUS       62056_10620        7980 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-mCherry-NAT1-Clox, complete sequence.
ACCESSION   MK652118
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7980)
  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 7980)
  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 7980)
  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..7980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             65..769
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     regulatory      773..1028
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     1038..1054
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1068..1101
                     /label=loxP
                     /note="loxP"
     terminator      complement(1512..1709)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1727..2290)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(2310..2653)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2717..4048
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(4076..4479,4644..5271)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59761.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4480..4643
                     /note="modified TUB2 intron sequence"
     3'UTR           5284..5414
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5415..5474
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5487..5520)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5618..5636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5894..6482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6656..7513)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7514..7618)
                     /label=AmpR promoter
     promoter        7964..7980
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggtgctg gcgcaggtgc
       61 ttctgtttca aaaggtgaag aagataatat ggctattatt aaagaattta tgagatttaa
      121 agttcatatg gaaggttcag ttaatggtca tgaatttgaa attgaaggtg aaggtgaagg
      181 tagaccatat gaaggtactc aaactgctaa attgaaagtt actaaaggtg gttcattacc
      241 atttgcttgg gatattttgt caccacaatt tatgtatggt tcaaaagctt atgttaaaca
      301 tccagctgat attccagatt atttaaaatt gtcatttcca gaaggtttta aatgggaaag
      361 agttatgaat tttgaagatg gtggtgttgt tactgttact caagattcat cattacaaga
      421 tggtgaattt atttataagg ttaaattgag aggtactaat tttccatcag atggtccagt
      481 tatgcaaaaa aaaactatgg gttgggaagc ttcatcagaa agaatgtatc cagaagatgg
      541 tgctttaaaa ggtgaaatta aacaaagatt gaaattaaaa gatggtggtc attatgatgc
      601 tgaagttaaa actacttata aagctaaaaa accagttcaa ttaccaggtg cttataatgt
      661 taatattaaa ttggatatta cttcacataa tgaagattat actattgttg aacaatatga
      721 aagagctgaa ggtagacatt caactggtgg tatggatgaa ttatataaat aaggcgcgcc
      781 attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca
      841 gttattaccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa aaaaaattgg
      901 aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc cagaaccaag
      961 taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa agatggacgc
     1021 atagatccgg cggccgctct agaactagtg gatccagatc ttgatccata acttcgtata
     1081 gcatacatta tacgaagtta tagatccagt actaatagga attgatttgg atggtataaa
     1141 cggaaacaaa aaaaagagct ggtactactt tctttaaaat tattttatta tttgatttta
     1201 tttaatagta tatattatat tttgaacgta gattattttg ttgaaagttg ctgtagtgcc
     1261 attgattcgt aacactaatt ctgtattagt cattcctctt gtttgatagt atccaaaaaa
     1321 acggctattt ttttgcaatc ttatttcctg catattatac agataacata atgaaagaaa
     1381 aaatcttttt ttttgttctt caatgatgat ttcaaccatt cttttaaaca ttgatcaatt
     1441 cctgagctca tcccattcat tccatcataa aatgtcgaca ctggatggcg gcgttagtat
     1501 cgaatcgaca gcagtatagc gaccagcatt cacatacgat tgacgcatga tattactttc
     1561 tgcgcactta acttcgcatc tgggcagatg atgtcgaggc gaaaaaaaat ataaatcacg
     1621 ctaacatttg attaaaatag aacaactaca atataaaaaa actatacaaa tgacaagttc
     1681 ttgaaaacaa gaatcttttt attgtcagta ctgactcgag ttattatgga catggcatag
     1741 acatatacaa agcttgttca ccatcggaag cagtaccatc gtataaagca gtatccaaac
     1801 cacacaaagt gaaacccatt cttctataag catgaatagc tggagcatta acattggtaa
     1861 cttccaacca caaatgacca gcacctcttt ctctggcgaa ttcagtagcc aaacccatca
     1921 aagctctacc aacaccatga cctctatgtt ctggagcaac ttcaatatct tcaacagtca
     1981 atcttctgtt ccaaccagaa taagaaacaa caacgaaacc agccaaatca ccatcatcac
     2041 cataagcaac gaaagttcta gaatctggat caccatcttc accagcatcg gattcatcat
     2101 cggattcatc atctgggaaa accttagtca atggtggatc aactggaact tctctcaaag
     2161 tgaaaccatc accagtagca gtaactctaa aaacagtatc ggtagtgaaa gaaccatcca
     2221 aagcttcaat agcttcagca tcacctggaa cagaagttct gtatctataa gcagtatcat
     2281 ccaaagtagt agacataatt gtaggatccg gttgtttatg ttcggatgtg atgtgagaac
     2341 tgtatcctag caagatttta aaaggaagta tatgaaagaa gaacctcagt ggcaaatcct
     2401 aaccttttat atttctctac aggggcgcgg cgtggggaca attcaacgcg tctgtgaggg
     2461 gagcgtttcc ctgctcgcag gtctgcagcg aggagccgta atttttgctt cgcgccgtgc
     2521 ggccatcaaa atgtatggat gcaaatgatt atacatgggg atgtatgggc taaatgtacg
     2581 ggcgacagtc acatcatgcc cctgagctgc gcacgtcaag actgtcaagg agggtattct
     2641 gggcctccat gtcgctggcc gggtgacccg gcggggacga ggcaagcttg atggaagttc
     2701 ctatactttc gctagcaatt gtctattcca agcctgtggc tttttttggt gattcacaat
     2761 acataaattg gcaaatacct cttctataga ctttaacctt tgatgaaatg gtatggataa
     2821 accacttttg tggtcagggt atatatattt ttattccaat ctggccttta taaattcaaa
     2881 tttccaaggg gactctgcta aaccttctga tataactata actctaataa tggctgtaaa
     2941 aagtgtgaaa aaaagatgat attcacaaaa aagataaagg ggatttcata ttgggtatct
     3001 agatttaaac atcgtatttc aacaatgaaa ctagaatcaa tacgaaatcc agcgcacaac
     3061 tgtgttcatt cccttgggaa ttatactgag taatgggaag aatgtgcaaa aaatacccgt
     3121 gtcttatata atttccattt gtgtagggct taaaagtgga caacttcagg ttacgaaacc
     3181 acgaaaatgt ctaaacgttt gatcaaattc gtaactcatg gaaaatcaca gcaacaaatt
     3241 aaggatggat atatccggta cttacaaata aaaacactaa agaagtataa attttgaact
     3301 gaaattctat agaaaatctt tttttttggg taaaattatg cgattgtggc tcatagtaac
     3361 gtaatcatac agttcaacaa acatcatttt gtcaaaggat ttctaatccc attctttttc
     3421 tacttggctt ttgagtatat atatatacat cctttgttca tcatcatgtt cgattgaatt
     3481 tttttcttgg ttgttattag attgtctact tgagtttttc ttggtgcaaa ttgtggtgtt
     3541 tttctaacaa atttttcaac atttaaagta tttctggaac ttttctacaa ggaacgacat
     3601 agatccatta atgcgcccct ctaaaacaat acccataatg tcttcttaag tatgcatctg
     3661 cttataaatg tttctggatt tactttagtt agaatgtcga cacgacaatg aactattgct
     3721 gtggatcacg tgcgatcact gaattatgag tttcttttta tttttctttt tgggcttttt
     3781 tttgcaattt tttttttcca cctttgctaa ttgcgtagtc acatataatg tatacaattt
     3841 ttgcaactct gacactataa ttgatccaat aataaagtat ataatagatc atcaagtata
     3901 cgtaatctcc ccaaaaaaaa attgcataaa ataaaatcta taaacctttt ttagattttt
     3961 agttgttcct tttttctttc tattcttcaa cttccaagtg ttgtcacttt ctccctgtta
     4021 actatattta aaagtaaata ccctcccccc cgggtatact cattaatcaa taattatgtc
     4081 taacttgttg accgttcacc aaaacttgcc agctttgcca gttgatgcta cctctgatga
     4141 agttagaaaa aacttgatgg atatgtttag agatagacaa gccttctctg aacacacctg
     4201 gaaaatgttg ttgtctgttt gtagatcctg ggctgcttgg tgtaaattga acaacagaaa
     4261 atggttccca gccgaaccag aagatgttag agattacttg ttgtacttgc aagctagagg
     4321 tttggctgtt aaaaccattc aacaacactt gggtcaattg aacatgttgc acagaagatc
     4381 cggtttgcca agaccatctg attctaacgc tgtttctttg gttatgagaa gaattagaaa
     4441 agaaaacgtt gatgctggtg aaagagctaa acaagccttg taggtataga cactgaagaa
     4501 aaaaaaattc tatcattgtt atgttgtgat ctttgatctt tagttgtcgg gttaacacct
     4561 gccaattgga tcaatacatc aatcaattaa ttctaatctt gaaaaaaaaa ttcattttac
     4621 taacaatttt ttcttttatt tagggctttc gaaagaaccg atttcgatca agttagatcc
     4681 ttgatggaaa actctgatag atgtcaagat attagaaact tggctttctt gggtattgct
     4741 tacaacacct tgttgagaat tgctgaaatt gctagaatta gagttaaaga tatttccaga
     4801 accgatggtg gtagaatgtt gattcacatt ggtagaacca aaaccttggt ttctaccgct
     4861 ggtgttgaaa aagccttgtc tttgggtgtt accaaattgg ttgaaagatg gatttctgtt
     4921 tctggtgttg ctgatgatcc aaacaactac ttgttctgta gagttagaaa aaacggtgtt
     4981 gctgctccat ctgctacctc tcaattatct accagagctt tggaaggtat tttcgaagct
     5041 acccacagat tgatttacgg tgctaaagat gattctggtc aaagatactt ggcttggtct
     5101 ggtcactctg ctagagttgg tgctgctaga gatatggcta gagccggcgt ttctattcca
     5161 gaaattatgc aagctggtgg ttggaccaac gttaacattg ttatgaacta cattagaaac
     5221 ttggattctg aaaccggtgc tatggttaga ttgttggaag atggtgatta atttatacct
     5281 aggtaagcaa atagctaaat tatatacgaa ttaatattat gattaagtgt ttacgtgagt
     5341 gcgatatttt tattactatc ttatacagtt gtatatactc tataaaatga gttgtctatt
     5401 aattaacgcg atgagtccct atttattttt ttatagttat gttagtatta agaacgttat
     5461 ttatatttca aattaattat ccatggataa cttcgtatag catacattat acgaagttat
     5521 tagggataac agggtaatct gcaggaaaaa cgagctcgaa ttcatcgatg atatcagatc
     5581 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     5641 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5701 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5761 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5821 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5881 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5941 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6001 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6061 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6121 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6181 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6241 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6301 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6361 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6421 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6601 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6661 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6721 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6781 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6841 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6901 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6961 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7021 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7081 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7141 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7201 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7261 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7321 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7381 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7441 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7501 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7561 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7621 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     7681 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     7741 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     7801 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     7861 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     7921 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
