LOCUS       62056_10645        7429 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-myc-NAT1-Clox, complete sequence.
ACCESSION   MK652128
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7429)
  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 7429)
  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 7429)
  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..7429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..83
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     regulatory      222..477
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     487..503
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    517..550
                     /label=loxP
                     /note="loxP"
     terminator      complement(961..1158)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1176..1739)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(1759..2102)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      2166..3497
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3525..3928,4093..4720)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59785.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3929..4092
                     /note="modified TUB2 intron sequence"
     3'UTR           4733..4863
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4864..4923
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae CYC1 gene"
                     /regulatory_class="terminator"
     protein_bind    complement(4936..4969)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(5067..5085)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(5343..5931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6105..6962)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6963..7067)
                     /label=AmpR promoter
     promoter        7413..7429
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc ggggaacaga
       61 agcttatatc cgaagaagac ctcggagagc aaaagctcat ttcagaagag gatctaggcg
      121 aacagaaact aatctcggag gaggacctcg gtgaacaaaa gcttatctct gaggaagatc
      181 ttggcgagca gaagctcata tcagaggaag acctagggta gggcgcgcca ttataagtaa
      241 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
      301 ggaatctcgg tcgtaatgat ttctataatg acgaaaaaaa aaaaattgga aagaaaaagc
      361 ttcatggcct ttataaaaag gaactatcca atacctcgcc agaaccaagt aacagtattt
      421 tacggggcac aaatcaagaa caataagaca ggactgtaaa gatggacgca tagatccggc
      481 ggccgctcta gaactagtgg atccagatct tgatccataa cttcgtatag catacattat
      541 acgaagttat agatccagta ctaataggaa ttgatttgga tggtataaac ggaaacaaaa
      601 aaaagagctg gtactacttt ctttaaaatt attttattat ttgattttat ttaatagtat
      661 atattatatt ttgaacgtag attattttgt tgaaagttgc tgtagtgcca ttgattcgta
      721 acactaattc tgtattagtc attcctcttg tttgatagta tccaaaaaaa cggctatttt
      781 tttgcaatct tatttcctgc atattataca gataacataa tgaaagaaaa aatctttttt
      841 tttgttcttc aatgatgatt tcaaccattc ttttaaacat tgatcaattc ctgagctcat
      901 cccattcatt ccatcataaa atgtcgacac tggatggcgg cgttagtatc gaatcgacag
      961 cagtatagcg accagcattc acatacgatt gacgcatgat attactttct gcgcacttaa
     1021 cttcgcatct gggcagatga tgtcgaggcg aaaaaaaata taaatcacgc taacatttga
     1081 ttaaaataga acaactacaa tataaaaaaa ctatacaaat gacaagttct tgaaaacaag
     1141 aatcttttta ttgtcagtac tgactcgagt tattatggac atggcataga catatacaaa
     1201 gcttgttcac catcggaagc agtaccatcg tataaagcag tatccaaacc acacaaagtg
     1261 aaacccattc ttctataagc atgaatagct ggagcattaa cattggtaac ttccaaccac
     1321 aaatgaccag cacctctttc tctggcgaat tcagtagcca aacccatcaa agctctacca
     1381 acaccatgac ctctatgttc tggagcaact tcaatatctt caacagtcaa tcttctgttc
     1441 caaccagaat aagaaacaac aacgaaacca gccaaatcac catcatcacc ataagcaacg
     1501 aaagttctag aatctggatc accatcttca ccagcatcgg attcatcatc ggattcatca
     1561 tctgggaaaa ccttagtcaa tggtggatca actggaactt ctctcaaagt gaaaccatca
     1621 ccagtagcag taactctaaa aacagtatcg gtagtgaaag aaccatccaa agcttcaata
     1681 gcttcagcat cacctggaac agaagttctg tatctataag cagtatcatc caaagtagta
     1741 gacataattg taggatccgg ttgtttatgt tcggatgtga tgtgagaact gtatcctagc
     1801 aagattttaa aaggaagtat atgaaagaag aacctcagtg gcaaatccta accttttata
     1861 tttctctaca ggggcgcggc gtggggacaa ttcaacgcgt ctgtgagggg agcgtttccc
     1921 tgctcgcagg tctgcagcga ggagccgtaa tttttgcttc gcgccgtgcg gccatcaaaa
     1981 tgtatggatg caaatgatta tacatgggga tgtatgggct aaatgtacgg gcgacagtca
     2041 catcatgccc ctgagctgcg cacgtcaaga ctgtcaagga gggtattctg ggcctccatg
     2101 tcgctggccg ggtgacccgg cggggacgag gcaagcttga tggaagttcc tatactttcg
     2161 ctagcaattg tctattccaa gcctgtggct ttttttggtg attcacaata cataaattgg
     2221 caaatacctc ttctatagac tttaaccttt gatgaaatgg tatggataaa ccacttttgt
     2281 ggtcagggta tatatatttt tattccaatc tggcctttat aaattcaaat ttccaagggg
     2341 actctgctaa accttctgat ataactataa ctctaataat ggctgtaaaa agtgtgaaaa
     2401 aaagatgata ttcacaaaaa agataaaggg gatttcatat tgggtatcta gatttaaaca
     2461 tcgtatttca acaatgaaac tagaatcaat acgaaatcca gcgcacaact gtgttcattc
     2521 ccttgggaat tatactgagt aatgggaaga atgtgcaaaa aatacccgtg tcttatataa
     2581 tttccatttg tgtagggctt aaaagtggac aacttcaggt tacgaaacca cgaaaatgtc
     2641 taaacgtttg atcaaattcg taactcatgg aaaatcacag caacaaatta aggatggata
     2701 tatccggtac ttacaaataa aaacactaaa gaagtataaa ttttgaactg aaattctata
     2761 gaaaatcttt ttttttgggt aaaattatgc gattgtggct catagtaacg taatcataca
     2821 gttcaacaaa catcattttg tcaaaggatt tctaatccca ttctttttct acttggcttt
     2881 tgagtatata tatatacatc ctttgttcat catcatgttc gattgaattt ttttcttggt
     2941 tgttattaga ttgtctactt gagtttttct tggtgcaaat tgtggtgttt ttctaacaaa
     3001 tttttcaaca tttaaagtat ttctggaact tttctacaag gaacgacata gatccattaa
     3061 tgcgcccctc taaaacaata cccataatgt cttcttaagt atgcatctgc ttataaatgt
     3121 ttctggattt actttagtta gaatgtcgac acgacaatga actattgctg tggatcacgt
     3181 gcgatcactg aattatgagt ttctttttat ttttcttttt gggctttttt ttgcaatttt
     3241 ttttttccac ctttgctaat tgcgtagtca catataatgt atacaatttt tgcaactctg
     3301 acactataat tgatccaata ataaagtata taatagatca tcaagtatac gtaatctccc
     3361 caaaaaaaaa ttgcataaaa taaaatctat aaaccttttt tagattttta gttgttcctt
     3421 ttttctttct attcttcaac ttccaagtgt tgtcactttc tccctgttaa ctatatttaa
     3481 aagtaaatac cctccccccc gggtatactc attaatcaat aattatgtct aacttgttga
     3541 ccgttcacca aaacttgcca gctttgccag ttgatgctac ctctgatgaa gttagaaaaa
     3601 acttgatgga tatgtttaga gatagacaag ccttctctga acacacctgg aaaatgttgt
     3661 tgtctgtttg tagatcctgg gctgcttggt gtaaattgaa caacagaaaa tggttcccag
     3721 ccgaaccaga agatgttaga gattacttgt tgtacttgca agctagaggt ttggctgtta
     3781 aaaccattca acaacacttg ggtcaattga acatgttgca cagaagatcc ggtttgccaa
     3841 gaccatctga ttctaacgct gtttctttgg ttatgagaag aattagaaaa gaaaacgttg
     3901 atgctggtga aagagctaaa caagccttgt aggtatagac actgaagaaa aaaaaattct
     3961 atcattgtta tgttgtgatc tttgatcttt agttgtcggg ttaacacctg ccaattggat
     4021 caatacatca atcaattaat tctaatcttg aaaaaaaaat tcattttact aacaattttt
     4081 tcttttattt agggctttcg aaagaaccga tttcgatcaa gttagatcct tgatggaaaa
     4141 ctctgataga tgtcaagata ttagaaactt ggctttcttg ggtattgctt acaacacctt
     4201 gttgagaatt gctgaaattg ctagaattag agttaaagat atttccagaa ccgatggtgg
     4261 tagaatgttg attcacattg gtagaaccaa aaccttggtt tctaccgctg gtgttgaaaa
     4321 agccttgtct ttgggtgtta ccaaattggt tgaaagatgg atttctgttt ctggtgttgc
     4381 tgatgatcca aacaactact tgttctgtag agttagaaaa aacggtgttg ctgctccatc
     4441 tgctacctct caattatcta ccagagcttt ggaaggtatt ttcgaagcta cccacagatt
     4501 gatttacggt gctaaagatg attctggtca aagatacttg gcttggtctg gtcactctgc
     4561 tagagttggt gctgctagag atatggctag agccggcgtt tctattccag aaattatgca
     4621 agctggtggt tggaccaacg ttaacattgt tatgaactac attagaaact tggattctga
     4681 aaccggtgct atggttagat tgttggaaga tggtgattaa tttataccta ggtaagcaaa
     4741 tagctaaatt atatacgaat taatattatg attaagtgtt tacgtgagtg cgatattttt
     4801 attactatct tatacagttg tatatactct ataaaatgag ttgtctatta attaacgcga
     4861 tgagtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     4921 attaattatc catggataac ttcgtatagc atacattata cgaagttatt agggataaca
     4981 gggtaatctg caggaaaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     5041 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     5101 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5161 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5221 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5281 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5341 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5401 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5461 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5521 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5581 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5641 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5701 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5761 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5821 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7081 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     7141 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     7201 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     7261 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     7321 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     7381 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
