LOCUS       62056_10530        9417 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-3xGFP-NAT1-Clox, complete sequence.
ACCESSION   MK652103
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9417)
  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 9417)
  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 9417)
  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. 12
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9417
                     /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"
     terminator      complement(2949..3146)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3164..3727)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(3747..4090)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      4154..5485
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(5513..5916,6081..6708)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59725.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          5917..6080
                     /note="modified TUB2 intron sequence"
     3'UTR           6721..6851
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      6852..6911
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(6924..6957)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(7055..7073)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(7331..7919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8093..8950)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8951..9055)
                     /label=AmpR promoter
     promoter        9401..9417
                     /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 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 gagctcatcc cattcattcc atcataaaat gtcgacactg gatggcggcg ttagtatcga
     2941 atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
     3001 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
     3061 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
     3121 aaaacaagaa tctttttatt gtcagtactg actcgagtta ttatggacat ggcatagaca
     3181 tatacaaagc ttgttcacca tcggaagcag taccatcgta taaagcagta tccaaaccac
     3241 acaaagtgaa acccattctt ctataagcat gaatagctgg agcattaaca ttggtaactt
     3301 ccaaccacaa atgaccagca cctctttctc tggcgaattc agtagccaaa cccatcaaag
     3361 ctctaccaac accatgacct ctatgttctg gagcaacttc aatatcttca acagtcaatc
     3421 ttctgttcca accagaataa gaaacaacaa cgaaaccagc caaatcacca tcatcaccat
     3481 aagcaacgaa agttctagaa tctggatcac catcttcacc agcatcggat tcatcatcgg
     3541 attcatcatc tgggaaaacc ttagtcaatg gtggatcaac tggaacttct ctcaaagtga
     3601 aaccatcacc agtagcagta actctaaaaa cagtatcggt agtgaaagaa ccatccaaag
     3661 cttcaatagc ttcagcatca cctggaacag aagttctgta tctataagca gtatcatcca
     3721 aagtagtaga cataattgta ggatccggtt gtttatgttc ggatgtgatg tgagaactgt
     3781 atcctagcaa gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac
     3841 cttttatatt tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag
     3901 cgtttccctg ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc
     3961 catcaaaatg tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc
     4021 gacagtcaca tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg
     4081 cctccatgtc gctggccggg tgacccggcg gggacgaggc aagcttgatg gaagttccta
     4141 tactttcgct agcaattgtc tattccaagc ctgtggcttt ttttggtgat tcacaataca
     4201 taaattggca aatacctctt ctatagactt taacctttga tgaaatggta tggataaacc
     4261 acttttgtgg tcagggtata tatattttta ttccaatctg gcctttataa attcaaattt
     4321 ccaaggggac tctgctaaac cttctgatat aactataact ctaataatgg ctgtaaaaag
     4381 tgtgaaaaaa agatgatatt cacaaaaaag ataaagggga tttcatattg ggtatctaga
     4441 tttaaacatc gtatttcaac aatgaaacta gaatcaatac gaaatccagc gcacaactgt
     4501 gttcattccc ttgggaatta tactgagtaa tgggaagaat gtgcaaaaaa tacccgtgtc
     4561 ttatataatt tccatttgtg tagggcttaa aagtggacaa cttcaggtta cgaaaccacg
     4621 aaaatgtcta aacgtttgat caaattcgta actcatggaa aatcacagca acaaattaag
     4681 gatggatata tccggtactt acaaataaaa acactaaaga agtataaatt ttgaactgaa
     4741 attctataga aaatcttttt ttttgggtaa aattatgcga ttgtggctca tagtaacgta
     4801 atcatacagt tcaacaaaca tcattttgtc aaaggatttc taatcccatt ctttttctac
     4861 ttggcttttg agtatatata tatacatcct ttgttcatca tcatgttcga ttgaattttt
     4921 ttcttggttg ttattagatt gtctacttga gtttttcttg gtgcaaattg tggtgttttt
     4981 ctaacaaatt tttcaacatt taaagtattt ctggaacttt tctacaagga acgacataga
     5041 tccattaatg cgcccctcta aaacaatacc cataatgtct tcttaagtat gcatctgctt
     5101 ataaatgttt ctggatttac tttagttaga atgtcgacac gacaatgaac tattgctgtg
     5161 gatcacgtgc gatcactgaa ttatgagttt ctttttattt ttctttttgg gctttttttt
     5221 gcaatttttt ttttccacct ttgctaattg cgtagtcaca tataatgtat acaatttttg
     5281 caactctgac actataattg atccaataat aaagtatata atagatcatc aagtatacgt
     5341 aatctcccca aaaaaaaatt gcataaaata aaatctataa acctttttta gatttttagt
     5401 tgttcctttt ttctttctat tcttcaactt ccaagtgttg tcactttctc cctgttaact
     5461 atatttaaaa gtaaataccc tcccccccgg gtatactcat taatcaataa ttatgtctaa
     5521 cttgttgacc gttcaccaaa acttgccagc tttgccagtt gatgctacct ctgatgaagt
     5581 tagaaaaaac ttgatggata tgtttagaga tagacaagcc ttctctgaac acacctggaa
     5641 aatgttgttg tctgtttgta gatcctgggc tgcttggtgt aaattgaaca acagaaaatg
     5701 gttcccagcc gaaccagaag atgttagaga ttacttgttg tacttgcaag ctagaggttt
     5761 ggctgttaaa accattcaac aacacttggg tcaattgaac atgttgcaca gaagatccgg
     5821 tttgccaaga ccatctgatt ctaacgctgt ttctttggtt atgagaagaa ttagaaaaga
     5881 aaacgttgat gctggtgaaa gagctaaaca agccttgtag gtatagacac tgaagaaaaa
     5941 aaaattctat cattgttatg ttgtgatctt tgatctttag ttgtcgggtt aacacctgcc
     6001 aattggatca atacatcaat caattaattc taatcttgaa aaaaaaattc attttactaa
     6061 caattttttc ttttatttag ggctttcgaa agaaccgatt tcgatcaagt tagatccttg
     6121 atggaaaact ctgatagatg tcaagatatt agaaacttgg ctttcttggg tattgcttac
     6181 aacaccttgt tgagaattgc tgaaattgct agaattagag ttaaagatat ttccagaacc
     6241 gatggtggta gaatgttgat tcacattggt agaaccaaaa ccttggtttc taccgctggt
     6301 gttgaaaaag ccttgtcttt gggtgttacc aaattggttg aaagatggat ttctgtttct
     6361 ggtgttgctg atgatccaaa caactacttg ttctgtagag ttagaaaaaa cggtgttgct
     6421 gctccatctg ctacctctca attatctacc agagctttgg aaggtatttt cgaagctacc
     6481 cacagattga tttacggtgc taaagatgat tctggtcaaa gatacttggc ttggtctggt
     6541 cactctgcta gagttggtgc tgctagagat atggctagag ccggcgtttc tattccagaa
     6601 attatgcaag ctggtggttg gaccaacgtt aacattgtta tgaactacat tagaaacttg
     6661 gattctgaaa ccggtgctat ggttagattg ttggaagatg gtgattaatt tatacctagg
     6721 taagcaaata gctaaattat atacgaatta atattatgat taagtgttta cgtgagtgcg
     6781 atatttttat tactatctta tacagttgta tatactctat aaaatgagtt gtctattaat
     6841 taacgcgatg agtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     6901 tatttcaaat taattatcca tggataactt cgtatagcat acattatacg aagttattag
     6961 ggataacagg gtaatctgca ggaaaaacga gctcgaattc atcgatgata tcagatccac
     7021 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     7081 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7141 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7501 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     7561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7741 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8221 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8401 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8521 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8581 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9061 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     9121 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     9181 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     9241 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     9301 cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg
     9361 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
