LOCUS       62056_10675        7801 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-TAP-NAT1-Clox, complete sequence.
ACCESSION   MK652133
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7801)
  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 7801)
  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 7801)
  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..7801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..146
                     /label=CBP
                     /note="calmodulin-binding peptide"
     CDS             174..194
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             225..398
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
     regulatory      594..849
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     859..875
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    889..922
                     /label=loxP
                     /note="loxP"
     terminator      complement(1333..1530)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1548..2111)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(2131..2474)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2538..3869
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3897..4300,4465..5092)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59797.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          4301..4464
                     /note="modified TUB2 intron sequence"
     3'UTR           5105..5235
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      5236..5295
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(5308..5341)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5439..5457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5715..6303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6477..7334)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7335..7439)
                     /label=AmpR promoter
     promoter        7785..7801
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 atccatggaa gagaagatgg aaaaagaatt tcatagccgt ctcagcagcc aaccgcttta
      121 agaaaatctc atcctccggg gcacttgatt atgatattcc aactactgct agcgagaatt
      181 tgtattttca gggagaattc ggccttgcgc aacacgatga agccgtggac aacaaattca
      241 acaaagaaca acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac
      301 aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag
      361 cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag
      421 aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa
      481 acgccttcat ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag
      541 ctaaaaagct aaatgatgct caggcgccga aagtagacgc gaatcatcag tgaggcgcgc
      601 cattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
      661 agttattacc cgggaatctc ggtcgtaatg atttctataa tgacgaaaaa aaaaaaattg
      721 gaaagaaaaa gcttcatggc ctttataaaa aggaactatc caatacctcg ccagaaccaa
      781 gtaacagtat tttacggggc acaaatcaag aacaataaga caggactgta aagatggacg
      841 catagatccg gcggccgctc tagaactagt ggatccagat cttgatccat aacttcgtat
      901 agcatacatt atacgaagtt atagatccag tactaatagg aattgatttg gatggtataa
      961 acggaaacaa aaaaaagagc tggtactact ttctttaaaa ttattttatt atttgatttt
     1021 atttaatagt atatattata ttttgaacgt agattatttt gttgaaagtt gctgtagtgc
     1081 cattgattcg taacactaat tctgtattag tcattcctct tgtttgatag tatccaaaaa
     1141 aacggctatt tttttgcaat cttatttcct gcatattata cagataacat aatgaaagaa
     1201 aaaatctttt tttttgttct tcaatgatga tttcaaccat tcttttaaac attgatcaat
     1261 tcctgagctc atcccattca ttccatcata aaatgtcgac actggatggc ggcgttagta
     1321 tcgaatcgac agcagtatag cgaccagcat tcacatacga ttgacgcatg atattacttt
     1381 ctgcgcactt aacttcgcat ctgggcagat gatgtcgagg cgaaaaaaaa tataaatcac
     1441 gctaacattt gattaaaata gaacaactac aatataaaaa aactatacaa atgacaagtt
     1501 cttgaaaaca agaatctttt tattgtcagt actgactcga gttattatgg acatggcata
     1561 gacatataca aagcttgttc accatcggaa gcagtaccat cgtataaagc agtatccaaa
     1621 ccacacaaag tgaaacccat tcttctataa gcatgaatag ctggagcatt aacattggta
     1681 acttccaacc acaaatgacc agcacctctt tctctggcga attcagtagc caaacccatc
     1741 aaagctctac caacaccatg acctctatgt tctggagcaa cttcaatatc ttcaacagtc
     1801 aatcttctgt tccaaccaga ataagaaaca acaacgaaac cagccaaatc accatcatca
     1861 ccataagcaa cgaaagttct agaatctgga tcaccatctt caccagcatc ggattcatca
     1921 tcggattcat catctgggaa aaccttagtc aatggtggat caactggaac ttctctcaaa
     1981 gtgaaaccat caccagtagc agtaactcta aaaacagtat cggtagtgaa agaaccatcc
     2041 aaagcttcaa tagcttcagc atcacctgga acagaagttc tgtatctata agcagtatca
     2101 tccaaagtag tagacataat tgtaggatcc ggttgtttat gttcggatgt gatgtgagaa
     2161 ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc
     2221 taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg
     2281 ggagcgtttc cctgctcgca ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg
     2341 cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac
     2401 gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc
     2461 tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagctt gatggaagtt
     2521 cctatacttt cgctagcaat tgtctattcc aagcctgtgg ctttttttgg tgattcacaa
     2581 tacataaatt ggcaaatacc tcttctatag actttaacct ttgatgaaat ggtatggata
     2641 aaccactttt gtggtcaggg tatatatatt tttattccaa tctggccttt ataaattcaa
     2701 atttccaagg ggactctgct aaaccttctg atataactat aactctaata atggctgtaa
     2761 aaagtgtgaa aaaaagatga tattcacaaa aaagataaag gggatttcat attgggtatc
     2821 tagatttaaa catcgtattt caacaatgaa actagaatca atacgaaatc cagcgcacaa
     2881 ctgtgttcat tcccttggga attatactga gtaatgggaa gaatgtgcaa aaaatacccg
     2941 tgtcttatat aatttccatt tgtgtagggc ttaaaagtgg acaacttcag gttacgaaac
     3001 cacgaaaatg tctaaacgtt tgatcaaatt cgtaactcat ggaaaatcac agcaacaaat
     3061 taaggatgga tatatccggt acttacaaat aaaaacacta aagaagtata aattttgaac
     3121 tgaaattcta tagaaaatct ttttttttgg gtaaaattat gcgattgtgg ctcatagtaa
     3181 cgtaatcata cagttcaaca aacatcattt tgtcaaagga tttctaatcc cattcttttt
     3241 ctacttggct tttgagtata tatatataca tcctttgttc atcatcatgt tcgattgaat
     3301 ttttttcttg gttgttatta gattgtctac ttgagttttt cttggtgcaa attgtggtgt
     3361 ttttctaaca aatttttcaa catttaaagt atttctggaa cttttctaca aggaacgaca
     3421 tagatccatt aatgcgcccc tctaaaacaa tacccataat gtcttcttaa gtatgcatct
     3481 gcttataaat gtttctggat ttactttagt tagaatgtcg acacgacaat gaactattgc
     3541 tgtggatcac gtgcgatcac tgaattatga gtttcttttt atttttcttt ttgggctttt
     3601 ttttgcaatt ttttttttcc acctttgcta attgcgtagt cacatataat gtatacaatt
     3661 tttgcaactc tgacactata attgatccaa taataaagta tataatagat catcaagtat
     3721 acgtaatctc cccaaaaaaa aattgcataa aataaaatct ataaaccttt tttagatttt
     3781 tagttgttcc ttttttcttt ctattcttca acttccaagt gttgtcactt tctccctgtt
     3841 aactatattt aaaagtaaat accctccccc ccgggtatac tcattaatca ataattatgt
     3901 ctaacttgtt gaccgttcac caaaacttgc cagctttgcc agttgatgct acctctgatg
     3961 aagttagaaa aaacttgatg gatatgttta gagatagaca agccttctct gaacacacct
     4021 ggaaaatgtt gttgtctgtt tgtagatcct gggctgcttg gtgtaaattg aacaacagaa
     4081 aatggttccc agccgaacca gaagatgtta gagattactt gttgtacttg caagctagag
     4141 gtttggctgt taaaaccatt caacaacact tgggtcaatt gaacatgttg cacagaagat
     4201 ccggtttgcc aagaccatct gattctaacg ctgtttcttt ggttatgaga agaattagaa
     4261 aagaaaacgt tgatgctggt gaaagagcta aacaagcctt gtaggtatag acactgaaga
     4321 aaaaaaaatt ctatcattgt tatgttgtga tctttgatct ttagttgtcg ggttaacacc
     4381 tgccaattgg atcaatacat caatcaatta attctaatct tgaaaaaaaa attcatttta
     4441 ctaacaattt tttcttttat ttagggcttt cgaaagaacc gatttcgatc aagttagatc
     4501 cttgatggaa aactctgata gatgtcaaga tattagaaac ttggctttct tgggtattgc
     4561 ttacaacacc ttgttgagaa ttgctgaaat tgctagaatt agagttaaag atatttccag
     4621 aaccgatggt ggtagaatgt tgattcacat tggtagaacc aaaaccttgg tttctaccgc
     4681 tggtgttgaa aaagccttgt ctttgggtgt taccaaattg gttgaaagat ggatttctgt
     4741 ttctggtgtt gctgatgatc caaacaacta cttgttctgt agagttagaa aaaacggtgt
     4801 tgctgctcca tctgctacct ctcaattatc taccagagct ttggaaggta ttttcgaagc
     4861 tacccacaga ttgatttacg gtgctaaaga tgattctggt caaagatact tggcttggtc
     4921 tggtcactct gctagagttg gtgctgctag agatatggct agagccggcg tttctattcc
     4981 agaaattatg caagctggtg gttggaccaa cgttaacatt gttatgaact acattagaaa
     5041 cttggattct gaaaccggtg ctatggttag attgttggaa gatggtgatt aatttatacc
     5101 taggtaagca aatagctaaa ttatatacga attaatatta tgattaagtg tttacgtgag
     5161 tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg agttgtctat
     5221 taattaacgc gatgagtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     5281 tttatatttc aaattaatta tccatggata acttcgtata gcatacatta tacgaagtta
     5341 ttagggataa cagggtaatc ttcaggaaaa acgagctcga attcatcgat gatatcagat
     5401 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     5461 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5521 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5881 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6121 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6361 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7441 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7501 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     7561 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     7621 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     7681 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
     7741 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     7801 a
//
