LOCUS       62056_10580        7359 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-HA-NAT1-Clox, complete sequence.
ACCESSION   MK652123
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  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 7359)
  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 7359)
  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..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             45..71
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             75..101
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             105..131
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     regulatory      135..390
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     400..416
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    430..463
                     /label=loxP
                     /note="loxP"
     terminator      complement(874..1071)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1089..1652)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(1672..2015)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2079..3410
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3438..3841,4006..4633)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59773.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3842..4005
                     /note="modified TUB2 intron sequence"
     3'UTR           4646..4776
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4777..4836
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(4849..4882)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4980..4998)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5256..5844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6018..6875)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6876..6980)
                     /label=AmpR promoter
     promoter        7326..7344
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaagcttcgt acgctgcagg tcgacgggtc gacggatccc cgggtaccca tacgatgttc
       61 ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatatcca tatgacgttc
      121 cagattacgc ttagggcgcg ccattataag taaatgcatg tatactaaac tcacaaatta
      181 gagcttcaat ttaattatat cagttattac ccgggaatct cggtcgtaat gatttctata
      241 atgacgaaaa aaaaaaaatt ggaaagaaaa agcttcatgg cctttataaa aaggaactat
      301 ccaatacctc gccagaacca agtaacagta ttttacgggg cacaaatcaa gaacaataag
      361 acaggactgt aaagatggac gcatagatcc ggcggccgct ctagaactag tggatccaga
      421 tcttgatcca taacttcgta tagcatacat tatacgaagt tatagatcca gtactaatag
      481 gaattgattt ggatggtata aacggaaaca aaaaaaagag ctggtactac tttctttaaa
      541 attattttat tatttgattt tatttaatag tatatattat attttgaacg tagattattt
      601 tgttgaaagt tgctgtagtg ccattgattc gtaacactaa ttctgtatta gtcattcctc
      661 ttgtttgata gtatccaaaa aaacggctat ttttttgcaa tcttatttcc tgcatattat
      721 acagataaca taatgaaaga aaaaatcttt ttttttgttc ttcaatgatg atttcaacca
      781 ttcttttaaa cattgatcaa ttcctgagct catcccattc attccatcat aaaatgtcga
      841 cactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca ttcacatacg
      901 attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga tgatgtcgag
      961 gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta caatataaaa
     1021 aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag tactgactcg
     1081 agttattatg gacatggcat agacatatac aaagcttgtt caccatcgga agcagtacca
     1141 tcgtataaag cagtatccaa accacacaaa gtgaaaccca ttcttctata agcatgaata
     1201 gctggagcat taacattggt aacttccaac cacaaatgac cagcacctct ttctctggcg
     1261 aattcagtag ccaaacccat caaagctcta ccaacaccat gacctctatg ttctggagca
     1321 acttcaatat cttcaacagt caatcttctg ttccaaccag aataagaaac aacaacgaaa
     1381 ccagccaaat caccatcatc accataagca acgaaagttc tagaatctgg atcaccatct
     1441 tcaccagcat cggattcatc atcggattca tcatctggga aaaccttagt caatggtgga
     1501 tcaactggaa cttctctcaa agtgaaacca tcaccagtag cagtaactct aaaaacagta
     1561 tcggtagtga aagaaccatc caaagcttca atagcttcag catcacctgg aacagaagtt
     1621 ctgtatctat aagcagtatc atccaaagta gtagacataa ttgtaggatc cggttgttta
     1681 tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag tatatgaaag
     1741 aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc ggcgtgggga
     1801 caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag cgaggagccg
     1861 taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga ttatacatgg
     1921 ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct gcgcacgtca
     1981 agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac
     2041 gaggcaagct tgatggaagt tcctatactt tcgctagcaa ttgtctattc caagcctgtg
     2101 gctttttttg gtgattcaca atacataaat tggcaaatac ctcttctata gactttaacc
     2161 tttgatgaaa tggtatggat aaaccacttt tgtggtcagg gtatatatat ttttattcca
     2221 atctggcctt tataaattca aatttccaag gggactctgc taaaccttct gatataacta
     2281 taactctaat aatggctgta aaaagtgtga aaaaaagatg atattcacaa aaaagataaa
     2341 ggggatttca tattgggtat ctagatttaa acatcgtatt tcaacaatga aactagaatc
     2401 aatacgaaat ccagcgcaca actgtgttca ttcccttggg aattatactg agtaatggga
     2461 agaatgtgca aaaaataccc gtgtcttata taatttccat ttgtgtaggg cttaaaagtg
     2521 gacaacttca ggttacgaaa ccacgaaaat gtctaaacgt ttgatcaaat tcgtaactca
     2581 tggaaaatca cagcaacaaa ttaaggatgg atatatccgg tacttacaaa taaaaacact
     2641 aaagaagtat aaattttgaa ctgaaattct atagaaaatc tttttttttg ggtaaaatta
     2701 tgcgattgtg gctcatagta acgtaatcat acagttcaac aaacatcatt ttgtcaaagg
     2761 atttctaatc ccattctttt tctacttggc ttttgagtat atatatatac atcctttgtt
     2821 catcatcatg ttcgattgaa tttttttctt ggttgttatt agattgtcta cttgagtttt
     2881 tcttggtgca aattgtggtg tttttctaac aaatttttca acatttaaag tatttctgga
     2941 acttttctac aaggaacgac atagatccat taatgcgccc ctctaaaaca atacccataa
     3001 tgtcttctta agtatgcatc tgcttataaa tgtttctgga tttactttag ttagaatgtc
     3061 gacacgacaa tgaactattg ctgtggatca cgtgcgatca ctgaattatg agtttctttt
     3121 tatttttctt tttgggcttt tttttgcaat tttttttttc cacctttgct aattgcgtag
     3181 tcacatataa tgtatacaat ttttgcaact ctgacactat aattgatcca ataataaagt
     3241 atataataga tcatcaagta tacgtaatct ccccaaaaaa aaattgcata aaataaaatc
     3301 tataaacctt ttttagattt ttagttgttc cttttttctt tctattcttc aacttccaag
     3361 tgttgtcact ttctccctgt taactatatt taaaagtaaa taccctcccc cccgggtata
     3421 ctcattaatc aataattatg tctaacttgt tgaccgttca ccaaaacttg ccagctttgc
     3481 cagttgatgc tacctctgat gaagttagaa aaaacttgat ggatatgttt agagatagac
     3541 aagccttctc tgaacacacc tggaaaatgt tgttgtctgt ttgtagatcc tgggctgctt
     3601 ggtgtaaatt gaacaacaga aaatggttcc cagccgaacc agaagatgtt agagattact
     3661 tgttgtactt gcaagctaga ggtttggctg ttaaaaccat tcaacaacac ttgggtcaat
     3721 tgaacatgtt gcacagaaga tccggtttgc caagaccatc tgattctaac gctgtttctt
     3781 tggttatgag aagaattaga aaagaaaacg ttgatgctgg tgaaagagct aaacaagcct
     3841 tgtaggtata gacactgaag aaaaaaaaat tctatcattg ttatgttgtg atctttgatc
     3901 tttagttgtc gggttaacac ctgccaattg gatcaataca tcaatcaatt aattctaatc
     3961 ttgaaaaaaa aattcatttt actaacaatt ttttctttta tttagggctt tcgaaagaac
     4021 cgatttcgat caagttagat ccttgatgga aaactctgat agatgtcaag atattagaaa
     4081 cttggctttc ttgggtattg cttacaacac cttgttgaga attgctgaaa ttgctagaat
     4141 tagagttaaa gatatttcca gaaccgatgg tggtagaatg ttgattcaca ttggtagaac
     4201 caaaaccttg gtttctaccg ctggtgttga aaaagccttg tctttgggtg ttaccaaatt
     4261 ggttgaaaga tggatttctg tttctggtgt tgctgatgat ccaaacaact acttgttctg
     4321 tagagttaga aaaaacggtg ttgctgctcc atctgctacc tctcaattat ctaccagagc
     4381 tttggaaggt attttcgaag ctacccacag attgatttac ggtgctaaag atgattctgg
     4441 tcaaagatac ttggcttggt ctggtcactc tgctagagtt ggtgctgcta gagatatggc
     4501 tagagccggc gtttctattc cagaaattat gcaagctggt ggttggacca acgttaacat
     4561 tgttatgaac tacattagaa acttggattc tgaaaccggt gctatggtta gattgttgga
     4621 agatggtgat taatttatac ctaggtaagc aaatagctaa attatatacg aattaatatt
     4681 atgattaagt gtttacgtga gtgcgatatt tttattacta tcttatacag ttgtatatac
     4741 tctataaaat gagttgtcta ttaattaacg cgatgagtcc ctatttattt ttttatagtt
     4801 atgttagtat taagaacgtt atttatattt caaattaatt atccatggat aacttcgtat
     4861 agcatacatt atacgaagtt attagggata acagggtaat ctgcaggaaa aacgagctcg
     4921 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     4981 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     5041 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5221 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5281 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5341 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5401 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     5461 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5521 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5581 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5641 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5701 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5821 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5881 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5941 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6001 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6061 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6121 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6181 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6241 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6301 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6361 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6421 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6481 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6541 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6601 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6661 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6721 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6781 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6841 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6901 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6961 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7021 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7081 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7141 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7201 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     7261 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     7321 atacgattta ggtgacacta tagaacgcgg ccgccagct
//
