LOCUS       62056_10555        7983 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-GFP-NAT1-Clox, complete sequence.
ACCESSION   MK652108
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  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 7983)
  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 7983)
  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..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             62..772
                     /label=GFP
                     /note="green fluorescent protein"
     regulatory      776..1031
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     1041..1057
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1071..1104
                     /label=loxP
                     /note="loxP"
     terminator      complement(1515..1712)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1730..2293)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(2313..2656)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2720..4051
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(4079..4482,4647..5274)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59737.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4483..4646
                     /note="modified TUB2 intron sequence"
     3'UTR           5287..5417
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5418..5477
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5490..5523)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5621..5639)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5897..6485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6659..7516)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7517..7621)
                     /label=AmpR promoter
     promoter        7967..7983
                     /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 aatagggcgc
      781 gccattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
      841 tcagttatta cccgggaatc tcggtcgtaa tgatttctat aatgacgaaa aaaaaaaaat
      901 tggaaagaaa aagcttcatg gcctttataa aaaggaacta tccaatacct cgccagaacc
      961 aagtaacagt attttacggg gcacaaatca agaacaataa gacaggactg taaagatgga
     1021 cgcatagatc cggcggccgc tctagaacta gtggatccag atcttgatcc ataacttcgt
     1081 atagcataca ttatacgaag ttatagatcc agtactaata ggaattgatt tggatggtat
     1141 aaacggaaac aaaaaaaaga gctggtacta ctttctttaa aattatttta ttatttgatt
     1201 ttatttaata gtatatatta tattttgaac gtagattatt ttgttgaaag ttgctgtagt
     1261 gccattgatt cgtaacacta attctgtatt agtcattcct cttgtttgat agtatccaaa
     1321 aaaacggcta tttttttgca atcttatttc ctgcatatta tacagataac ataatgaaag
     1381 aaaaaatctt tttttttgtt cttcaatgat gatttcaacc attcttttaa acattgatca
     1441 attcctgagc tcatcccatt cattccatca taaaatgtcg acactggatg gcggcgttag
     1501 tatcgaatcg acagcagtat agcgaccagc attcacatac gattgacgca tgatattact
     1561 ttctgcgcac ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa aatataaatc
     1621 acgctaacat ttgattaaaa tagaacaact acaatataaa aaaactatac aaatgacaag
     1681 ttcttgaaaa caagaatctt tttattgtca gtactgactc gagttattat ggacatggca
     1741 tagacatata caaagcttgt tcaccatcgg aagcagtacc atcgtataaa gcagtatcca
     1801 aaccacacaa agtgaaaccc attcttctat aagcatgaat agctggagca ttaacattgg
     1861 taacttccaa ccacaaatga ccagcacctc tttctctggc gaattcagta gccaaaccca
     1921 tcaaagctct accaacacca tgacctctat gttctggagc aacttcaata tcttcaacag
     1981 tcaatcttct gttccaacca gaataagaaa caacaacgaa accagccaaa tcaccatcat
     2041 caccataagc aacgaaagtt ctagaatctg gatcaccatc ttcaccagca tcggattcat
     2101 catcggattc atcatctggg aaaaccttag tcaatggtgg atcaactgga acttctctca
     2161 aagtgaaacc atcaccagta gcagtaactc taaaaacagt atcggtagtg aaagaaccat
     2221 ccaaagcttc aatagcttca gcatcacctg gaacagaagt tctgtatcta taagcagtat
     2281 catccaaagt agtagacata attgtaggat ccggttgttt atgttcggat gtgatgtgag
     2341 aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat
     2401 cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac gcgtctgtga
     2461 ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg cttcgcgccg
     2521 tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg ggctaaatgt
     2581 acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca aggagggtat
     2641 tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc ttgatggaag
     2701 ttcctatact ttcgctagca attgtctatt ccaagcctgt ggcttttttt ggtgattcac
     2761 aatacataaa ttggcaaata cctcttctat agactttaac ctttgatgaa atggtatgga
     2821 taaaccactt ttgtggtcag ggtatatata tttttattcc aatctggcct ttataaattc
     2881 aaatttccaa ggggactctg ctaaaccttc tgatataact ataactctaa taatggctgt
     2941 aaaaagtgtg aaaaaaagat gatattcaca aaaaagataa aggggatttc atattgggta
     3001 tctagattta aacatcgtat ttcaacaatg aaactagaat caatacgaaa tccagcgcac
     3061 aactgtgttc attcccttgg gaattatact gagtaatggg aagaatgtgc aaaaaatacc
     3121 cgtgtcttat ataatttcca tttgtgtagg gcttaaaagt ggacaacttc aggttacgaa
     3181 accacgaaaa tgtctaaacg tttgatcaaa ttcgtaactc atggaaaatc acagcaacaa
     3241 attaaggatg gatatatccg gtacttacaa ataaaaacac taaagaagta taaattttga
     3301 actgaaattc tatagaaaat cttttttttt gggtaaaatt atgcgattgt ggctcatagt
     3361 aacgtaatca tacagttcaa caaacatcat tttgtcaaag gatttctaat cccattcttt
     3421 ttctacttgg cttttgagta tatatatata catcctttgt tcatcatcat gttcgattga
     3481 atttttttct tggttgttat tagattgtct acttgagttt ttcttggtgc aaattgtggt
     3541 gtttttctaa caaatttttc aacatttaaa gtatttctgg aacttttcta caaggaacga
     3601 catagatcca ttaatgcgcc cctctaaaac aatacccata atgtcttctt aagtatgcat
     3661 ctgcttataa atgtttctgg atttacttta gttagaatgt cgacacgaca atgaactatt
     3721 gctgtggatc acgtgcgatc actgaattat gagtttcttt ttatttttct ttttgggctt
     3781 ttttttgcaa tttttttttt ccacctttgc taattgcgta gtcacatata atgtatacaa
     3841 tttttgcaac tctgacacta taattgatcc aataataaag tatataatag atcatcaagt
     3901 atacgtaatc tccccaaaaa aaaattgcat aaaataaaat ctataaacct tttttagatt
     3961 tttagttgtt ccttttttct ttctattctt caacttccaa gtgttgtcac tttctccctg
     4021 ttaactatat ttaaaagtaa ataccctccc ccccgggtat actcattaat caataattat
     4081 gtctaacttg ttgaccgttc accaaaactt gccagctttg ccagttgatg ctacctctga
     4141 tgaagttaga aaaaacttga tggatatgtt tagagataga caagccttct ctgaacacac
     4201 ctggaaaatg ttgttgtctg tttgtagatc ctgggctgct tggtgtaaat tgaacaacag
     4261 aaaatggttc ccagccgaac cagaagatgt tagagattac ttgttgtact tgcaagctag
     4321 aggtttggct gttaaaacca ttcaacaaca cttgggtcaa ttgaacatgt tgcacagaag
     4381 atccggtttg ccaagaccat ctgattctaa cgctgtttct ttggttatga gaagaattag
     4441 aaaagaaaac gttgatgctg gtgaaagagc taaacaagcc ttgtaggtat agacactgaa
     4501 gaaaaaaaaa ttctatcatt gttatgttgt gatctttgat ctttagttgt cgggttaaca
     4561 cctgccaatt ggatcaatac atcaatcaat taattctaat cttgaaaaaa aaattcattt
     4621 tactaacaat tttttctttt atttagggct ttcgaaagaa ccgatttcga tcaagttaga
     4681 tccttgatgg aaaactctga tagatgtcaa gatattagaa acttggcttt cttgggtatt
     4741 gcttacaaca ccttgttgag aattgctgaa attgctagaa ttagagttaa agatatttcc
     4801 agaaccgatg gtggtagaat gttgattcac attggtagaa ccaaaacctt ggtttctacc
     4861 gctggtgttg aaaaagcctt gtctttgggt gttaccaaat tggttgaaag atggatttct
     4921 gtttctggtg ttgctgatga tccaaacaac tacttgttct gtagagttag aaaaaacggt
     4981 gttgctgctc catctgctac ctctcaatta tctaccagag ctttggaagg tattttcgaa
     5041 gctacccaca gattgattta cggtgctaaa gatgattctg gtcaaagata cttggcttgg
     5101 tctggtcact ctgctagagt tggtgctgct agagatatgg ctagagccgg cgtttctatt
     5161 ccagaaatta tgcaagctgg tggttggacc aacgttaaca ttgttatgaa ctacattaga
     5221 aacttggatt ctgaaaccgg tgctatggtt agattgttgg aagatggtga ttaatttata
     5281 cctaggtaag caaatagcta aattatatac gaattaatat tatgattaag tgtttacgtg
     5341 agtgcgatat ttttattact atcttataca gttgtatata ctctataaaa tgagttgtct
     5401 attaattaac gcgatgagtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     5461 tatttatatt tcaaattaat tatccatgga taacttcgta tagcatacat tatacgaagt
     5521 tattagggat aacagggtaa tctgcaggaa aaacgagctc gaattcatcg atgatatcag
     5581 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     5641 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5701 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6061 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6301 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6421 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7081 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7621 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     7681 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     7741 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7801 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     7861 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     7921 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     7981 ata
//
