LOCUS       62056_10655        6995 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-NAT1-Clox, complete sequence.
ACCESSION   MK652113
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6995)
  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 6995)
  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 6995)
  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..6995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..69
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    83..116
                     /label=loxP
                     /note="loxP"
     terminator      complement(527..724)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(742..1305)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(1325..1668)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     regulatory      1732..3063
                     /note="CaMET3 promoter; inducible upon growth without
                     methionine repressible via addition of methionine and 
                     cysteine to the medium"
                     /regulatory_class="promoter"
     CDS             join(3091..3494,3659..4286)
                     /codon_start=1
                     /transl_table=11
                     /product="Cre"
                     /label=Cre
                     /note="Cre recombinase"
                     /protein_id="QDK59749.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          3495..3658
                     /note="modified TUB2 intron sequence"
     3'UTR           4299..4429
                     /label=Saccharomyces cerevisiae ADH1 3'UTR region
                     /note="Saccharomyces cerevisiae ADH1 3'UTR region"
     regulatory      4430..4489
                     /label=derived from Saccharomyces cerevisiae CYC1
                     /note="derived from Saccharomyces cerevisiae CYC1"
                     /regulatory_class="terminator"
     protein_bind    complement(4502..4535)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4633..4651)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(4909..5497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5671..6528)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6529..6633)
                     /label=AmpR promoter
     promoter        6979..6995
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggcggcc gctctagaac
       61 tagtggatcc agatcttgat ccataacttc gtatagcata cattatacga agttatagat
      121 ccagtactaa taggaattga tttggatggt ataaacggaa acaaaaaaaa gagctggtac
      181 tactttcttt aaaattattt tattatttga ttttatttaa tagtatatat tatattttga
      241 acgtagatta ttttgttgaa agttgctgta gtgccattga ttcgtaacac taattctgta
      301 ttagtcattc ctcttgtttg atagtatcca aaaaaacggc tatttttttg caatcttatt
      361 tcctgcatat tatacagata acataatgaa agaaaaaatc tttttttttg ttcttcaatg
      421 atgatttcaa ccattctttt aaacattgat caattcctga gctcatccca ttcattccat
      481 cataaaatgt cgacactgga tggcggcgtt agtatcgaat cgacagcagt atagcgacca
      541 gcattcacat acgattgacg catgatatta ctttctgcgc acttaacttc gcatctgggc
      601 agatgatgtc gaggcgaaaa aaaatataaa tcacgctaac atttgattaa aatagaacaa
      661 ctacaatata aaaaaactat acaaatgaca agttcttgaa aacaagaatc tttttattgt
      721 cagtactgac tcgagttatt atggacatgg catagacata tacaaagctt gttcaccatc
      781 ggaagcagta ccatcgtata aagcagtatc caaaccacac aaagtgaaac ccattcttct
      841 ataagcatga atagctggag cattaacatt ggtaacttcc aaccacaaat gaccagcacc
      901 tctttctctg gcgaattcag tagccaaacc catcaaagct ctaccaacac catgacctct
      961 atgttctgga gcaacttcaa tatcttcaac agtcaatctt ctgttccaac cagaataaga
     1021 aacaacaacg aaaccagcca aatcaccatc atcaccataa gcaacgaaag ttctagaatc
     1081 tggatcacca tcttcaccag catcggattc atcatcggat tcatcatctg ggaaaacctt
     1141 agtcaatggt ggatcaactg gaacttctct caaagtgaaa ccatcaccag tagcagtaac
     1201 tctaaaaaca gtatcggtag tgaaagaacc atccaaagct tcaatagctt cagcatcacc
     1261 tggaacagaa gttctgtatc tataagcagt atcatccaaa gtagtagaca taattgtagg
     1321 atccggttgt ttatgttcgg atgtgatgtg agaactgtat cctagcaaga ttttaaaagg
     1381 aagtatatga aagaagaacc tcagtggcaa atcctaacct tttatatttc tctacagggg
     1441 cgcggcgtgg ggacaattca acgcgtctgt gaggggagcg tttccctgct cgcaggtctg
     1501 cagcgaggag ccgtaatttt tgcttcgcgc cgtgcggcca tcaaaatgta tggatgcaaa
     1561 tgattataca tggggatgta tgggctaaat gtacgggcga cagtcacatc atgcccctga
     1621 gctgcgcacg tcaagactgt caaggagggt attctgggcc tccatgtcgc tggccgggtg
     1681 acccggcggg gacgaggcaa gcttgatgga agttcctata ctttcgctag caattgtcta
     1741 ttccaagcct gtggcttttt ttggtgattc acaatacata aattggcaaa tacctcttct
     1801 atagacttta acctttgatg aaatggtatg gataaaccac ttttgtggtc agggtatata
     1861 tatttttatt ccaatctggc ctttataaat tcaaatttcc aaggggactc tgctaaacct
     1921 tctgatataa ctataactct aataatggct gtaaaaagtg tgaaaaaaag atgatattca
     1981 caaaaaagat aaaggggatt tcatattggg tatctagatt taaacatcgt atttcaacaa
     2041 tgaaactaga atcaatacga aatccagcgc acaactgtgt tcattccctt gggaattata
     2101 ctgagtaatg ggaagaatgt gcaaaaaata cccgtgtctt atataatttc catttgtgta
     2161 gggcttaaaa gtggacaact tcaggttacg aaaccacgaa aatgtctaaa cgtttgatca
     2221 aattcgtaac tcatggaaaa tcacagcaac aaattaagga tggatatatc cggtacttac
     2281 aaataaaaac actaaagaag tataaatttt gaactgaaat tctatagaaa atcttttttt
     2341 ttgggtaaaa ttatgcgatt gtggctcata gtaacgtaat catacagttc aacaaacatc
     2401 attttgtcaa aggatttcta atcccattct ttttctactt ggcttttgag tatatatata
     2461 tacatccttt gttcatcatc atgttcgatt gaattttttt cttggttgtt attagattgt
     2521 ctacttgagt ttttcttggt gcaaattgtg gtgtttttct aacaaatttt tcaacattta
     2581 aagtatttct ggaacttttc tacaaggaac gacatagatc cattaatgcg cccctctaaa
     2641 acaataccca taatgtcttc ttaagtatgc atctgcttat aaatgtttct ggatttactt
     2701 tagttagaat gtcgacacga caatgaacta ttgctgtgga tcacgtgcga tcactgaatt
     2761 atgagtttct ttttattttt ctttttgggc ttttttttgc aatttttttt ttccaccttt
     2821 gctaattgcg tagtcacata taatgtatac aatttttgca actctgacac tataattgat
     2881 ccaataataa agtatataat agatcatcaa gtatacgtaa tctccccaaa aaaaaattgc
     2941 ataaaataaa atctataaac cttttttaga tttttagttg ttcctttttt ctttctattc
     3001 ttcaacttcc aagtgttgtc actttctccc tgttaactat atttaaaagt aaataccctc
     3061 ccccccgggt atactcatta atcaataatt atgtctaact tgttgaccgt tcaccaaaac
     3121 ttgccagctt tgccagttga tgctacctct gatgaagtta gaaaaaactt gatggatatg
     3181 tttagagata gacaagcctt ctctgaacac acctggaaaa tgttgttgtc tgtttgtaga
     3241 tcctgggctg cttggtgtaa attgaacaac agaaaatggt tcccagccga accagaagat
     3301 gttagagatt acttgttgta cttgcaagct agaggtttgg ctgttaaaac cattcaacaa
     3361 cacttgggtc aattgaacat gttgcacaga agatccggtt tgccaagacc atctgattct
     3421 aacgctgttt ctttggttat gagaagaatt agaaaagaaa acgttgatgc tggtgaaaga
     3481 gctaaacaag ccttgtaggt atagacactg aagaaaaaaa aattctatca ttgttatgtt
     3541 gtgatctttg atctttagtt gtcgggttaa cacctgccaa ttggatcaat acatcaatca
     3601 attaattcta atcttgaaaa aaaaattcat tttactaaca attttttctt ttatttaggg
     3661 ctttcgaaag aaccgatttc gatcaagtta gatccttgat ggaaaactct gatagatgtc
     3721 aagatattag aaacttggct ttcttgggta ttgcttacaa caccttgttg agaattgctg
     3781 aaattgctag aattagagtt aaagatattt ccagaaccga tggtggtaga atgttgattc
     3841 acattggtag aaccaaaacc ttggtttcta ccgctggtgt tgaaaaagcc ttgtctttgg
     3901 gtgttaccaa attggttgaa agatggattt ctgtttctgg tgttgctgat gatccaaaca
     3961 actacttgtt ctgtagagtt agaaaaaacg gtgttgctgc tccatctgct acctctcaat
     4021 tatctaccag agctttggaa ggtattttcg aagctaccca cagattgatt tacggtgcta
     4081 aagatgattc tggtcaaaga tacttggctt ggtctggtca ctctgctaga gttggtgctg
     4141 ctagagatat ggctagagcc ggcgtttcta ttccagaaat tatgcaagct ggtggttgga
     4201 ccaacgttaa cattgttatg aactacatta gaaacttgga ttctgaaacc ggtgctatgg
     4261 ttagattgtt ggaagatggt gattaattta tacctaggta agcaaatagc taaattatat
     4321 acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta ctatcttata
     4381 cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgag tccctattta
     4441 tttttttata gttatgttag tattaagaac gttatttata tttcaaatta attatccatg
     4501 gataacttcg tatagcatac attatacgaa gttattaggg ataacagggt aatctgcagg
     4561 aaaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg cggccgcgga
     4621 tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta
     4681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5101 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5341 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6541 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6601 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6661 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6721 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6781 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6841 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6901 gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     6961 atgtatcata cacatacgat ttaggtgaca ctata
//
