LOCUS       62056_9165        8545 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM018, complete sequence.
ACCESSION   MK431396
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8545)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz-Castro S, Johnson AD, Bennett 
            RJ.
  TITLE     Genetic Modification of Closely Related Candida Species
  JOURNAL   Front Microbiol 10, 357 (2019)
  PUBMED    30941104
REFERENCE   2  (bases 1 to 8545)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2019) Departmento de Ingenieria Genetica, Centro 
            de Investigacion y de Estudios Avanzados del Instituto Politecnico 
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera 
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico
REFERENCE   3  (bases 1 to 8545)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol 10, 357 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-JAN-2019) Departmento de Ingenieria Genetica, Centro de 
            Investigacion y de Estudios Avanzados del Instituto Politecnico 
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera 
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico"
FEATURES             Location/Qualifiers
     source          1..8545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..108
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     misc_recomb     623..656
                     /label=FLP recombination target sequence (FRT)
                     /note="FLP recombination target sequence (FRT)"
     misc_feature    663..1649
                     /label=promoter region from Candida tropicalis PCK1
                     /note="promoter region from Candida tropicalis PCK1"
     CDS             1650..2918
                     /label=FLP
                     /note="site-specific recombinase"
     misc_feature    2922..3309
                     /label=terminator region from Candida albicans ACT1
                     /note="terminator region from Candida albicans ACT1"
     misc_feature    3316..3813
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            3814..5040
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(3814..3823,4478..5040)
                     /codon_start=1
                     /transl_table=11
                     /gene="SAT1"
                     /product="streptomycin acetyltransferase"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="QBY25792.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     misc_feature    5044..5173
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(5178..5211)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(5213..5229)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5266..5284)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5305..5321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5329..5345)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5353..5383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5398..5419)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6515..6617
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6618..7274
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7765..7870)
                     /label=AmpR promoter
     rep_origin      7897..8352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8493..8509
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8519..8537
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggaagttcct atactttcta gagaatagga acttcggatc
       61 cccgggttaa ttaacggtga acaaaagcta atctccgagg aagacttgaa cggtgaacaa
      121 aaattaatct cagaagaaga cttgaacgga ctcgacggtg aacaaaagtt gatttctgaa
      181 gaagatttga acggtgaaca aaagctaatc tccgaggaag acttgaacgg tgaacaaaaa
      241 ttaatctcag aagaagactt gaacggactc gacggtgaac aaaagttgat ttctgaagaa
      301 gatttgaacg gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta
      361 atctcagaag aagacttgaa cggactcgac ggtgaacaaa agttgatttc tgaagaagat
      421 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
      481 tcagaagaag acttgaacgg actcgacggt gaacaaaagt tgatttctga agaagatttg
      541 aacggtgaac aaaagctaat ctccgaggaa gacttgaacg gtgaacaaaa attaatcaat
      601 cactagtgaa ttcgcgctcg aggaagttcc tatactttct agagaatagg aacttcggat
      661 ccgtgtgtgt attctgtata atttgtaaac tttattaatg tggttttttt gcttttaaag
      721 tattttgatt acgaaatgac gcgaaatttc catgaaaaaa aaaagaagaa agtttggggg
      781 ttcaattttg tacgataaca gtgttttttc actttttcca attcccctcc cccgggaatg
      841 aagtgacaaa cagttgttat taataactat atatccgaat taaacactat ttcctgccag
      901 gtgatgaata tactttatgg ttaccaagtt tcaaaaagga atattggtgg gttagtcggt
      961 ttacaagttt gaaaatcgat aataatcaag taaaaattat ctttcaaaat cagcaagaga
     1021 agaatatccc ctaaaattca ataaacaatt gttaaataaa caggcaattc taggcaaaat
     1081 gcagaatgat gttaaaaggt ttatgccgcc tttaactcat actcacatga cctttttctt
     1141 ctgataattt aattcaacta aactaagcaa aaaaaaatta aagcacagga aaggagttaa
     1201 ttaatttttc taaacaaaac atgttattgt gaataataca ctcaacatat tcattgattg
     1261 gctggcatgc ccagcatcaa agaaagaaat aaaccaaaag aaactccaga gtgaaaaatt
     1321 aatgtgcact aaaatattaa aatccacctc cttttccccg cgcgcacaca aagcgtagag
     1381 ctaactaaga gaaaaaaaca ttcgccttta ccccgggcta acgtttgtgc cggtaggtat
     1441 cgggatcatg ttcctaataa ttcctatgca tggtattttt accccagatt tcaaaagcac
     1501 ataaaagcac ttccggagaa aagggatagc tgctggcccc tataaaagtg gacgaaaatt
     1561 ccttccaaaa aaaatatttt gttctccttc ttttcttcct ttttatcatc attaacttaa
     1621 taacaaccaa gtttaaataa ttattcaata tgccacaatt tgatatatta tgtaaaacac
     1681 cacctaaggt gcttgttcgt cagtttgtgg aaaggtttga aagaccttca ggtgagaaaa
     1741 tagcattatg tgctgctgaa ctaacctatt tatgttggat gattacacat aacggaacag
     1801 caatcaagag agccacattc atgagctata atactatcat aagcaattcg ttgagtttcg
     1861 atattgtcaa taaatcactc cagtttaaat acaagacgca aaaagcaaca attttggaag
     1921 cctcattaaa gaaattgatt cctgcttggg aatttacaat tattccttac tatggacaaa
     1981 aacatcaatc tgatatcact gatattgtaa gtagtttgca attacagttc gaatcatcgg
     2041 aagaagcaga taagggaaat agccacagta aaaaaatgct taaagcactt ctaagtgagg
     2101 gtgaaagcat ctgggagatc actgagaaaa tactaaattc gtttgagtat acttcgagat
     2161 ttacaaaaac aaaaacttta taccaattcc tcttcctagc tactttcatc aattgtggaa
     2221 gattcagcga tattaagaac gttgatccga aatcatttaa attagtccaa aataagtatt
     2281 tgggagtaat aatccagtgt ttagtgacag agacaaagac aagcgttagt aggcacatat
     2341 acttctttag cgcaaggggt aggatcgatc cacttgtata tttggatgaa tttttgagga
     2401 attctgaacc agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc aataaacagg
     2461 aataccaatt attaaaagat aacttagtca gatcgtacaa taaagctttg aagaaaaatg
     2521 cgccttattc aatctttgct ataaaaaatg gcccaaaatc tcacattgga agacatttga
     2581 tgacctcatt tctttcaatg aagggcctaa cggagttgac taatgttgtg ggaaattgga
     2641 gcgataagcg tgcttctgcc gtggccagga caacgtatac tcatcagata acagcaatac
     2701 ctgatcacta cttcgcacta gtttctcggt actatgcata tgatccaata tcaaaggaaa
     2761 tgatagcatt gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa
     2821 agggtagtgc tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg
     2881 tactagacta cctttcatcc tacataaata gacgcatata agagtgaaat tctggaaatc
     2941 tggaaatctg gttttgtatt cttgttattc ttctttttgt tattacatat ataacttgtt
     3001 acttttttaa aaaaatcttt gtatatttta taaatatata aaactaaatt taagaaaaag
     3061 agaaaaatgt tttatttgag agattgaaat tttacttgaa tttagcttag cttttataaa
     3121 gtattattat gtaaaaaaac aaaacaaata tacattaaaa agttaagact ataaaatagc
     3181 cacccaaggc atttctatat cttgttgttg ttgttttcat cttctgtatc agaggaactt
     3241 attttattat tttcgtcacg ggtattttct cttgtttgat gattcatccc attcattcca
     3301 tcataaaatg tcgagcgtca aaactagaga ataataaaga aaacgatctt ttcaaaaaga
     3361 aaaaaccttt tagttttcct ttgttgttgt tgtgggtgtg tgctatttat attatatagt
     3421 ttactcataa taccataaaa tattcggttt gattaggtta ttttaataag ctaatttgtt
     3481 tctaatcgtg taatttatgc tgtgtatatt aagtagtgtg tgcactgccc aaaaatgttt
     3541 gttgtttata gtcggttaaa gagaaaaaag aaaaaaagat ccatacacac acgttaatta
     3601 gttgttcaac gtaatacact catattttgt tcttatttgc tttcggtcgc tgttctcacc
     3661 aagatttatt gccaacgaaa caattttttt ttatatattt tcagattttt ctttttttcc
     3721 tttcctttcc ttttctaatt ttcactcctg gttttctttc tttcttagaa acattatctc
     3781 gatattaata ttaaaaaaat ataatcattc aaaatggacg gtggtatgtt ttagtttagc
     3841 ttcaattcta attgattgat taatcagttg attggtttca atatgacaaa tgggtagggt
     3901 gggaaaactt cattttcaat tcagatcaaa cttttttgtt gtcgacataa tatttctcgt
     3961 ttgggatgtt actgtcacat taataataca cacacatcag cttataattt tgaaagtaat
     4021 ttatcagata tgttgtgacg atcaatggaa atggctaact tcaatgtatc tgttcttccc
     4081 ctttttcaaa gttcacgttt tttgattgat tgattgatct gtcggcagtg gtttcaaaac
     4141 cattcggtga gtaatcctat caatcaatgt tacgacaaaa ggctcaatat tcaaaattgc
     4201 aatgttttat gttttcctac gtgtacttgt gcaaggcaat tgattcaaca ttgcttttgg
     4261 tgtttgacga gtttctagtt tggacttgtg ttgttatctg ggctatacag atttcccggc
     4321 tcactatgaa tttttttttt cgacgctcag tgcacacaac tataaacaac acaaacacaa
     4381 acacagcaag aaaaaaaaaa aacgaacatt gaattgaaac caagccaact gaaaaattcc
     4441 ttatttaaat gactgtcata ctaacccatt tttatagaag aagttgctgc tttagttatc
     4501 gataacggtt ctcatatgaa aatttcggtg atccctgagc aggtggcgga aacattggat
     4561 gctgagaacc atttcattgt tcgtgaagtg ttcgatgtgc acctatccga ccaaggcttt
     4621 gaactatcta ccagaagtgt gagcccctac cggaaggatt acatctcgga tgatgactct
     4681 gatgaagact ctgcttgcta tggcgcattc atcgaccaag agcttgtcgg gaagattgaa
     4741 ctcaactcaa catggaacga tctagcctct atcgaacaca ttgttgtgtc gcacacgcac
     4801 cgaggcaaag gagtcgcgca cagtctcatc gaatttgcga aaaagtgggc actaagcaga
     4861 cagctccttg gcatacgatt agagacacaa acgaacaatg tacctgcctg caatttgtac
     4921 gcaaaatgtg gctttactct cggcggcatt gacctcttca cgtataaaac tagacctcaa
     4981 gtctcgaacg aaacagcgat gtactggtac tggttctcgg gagcacagga tgacgcctaa
     5041 catatgtgaa gtgtgaaggg ggagattttc actttattag atttgtatat atgtataata
     5101 aataaataaa taagttaaat aaataattag ataagggtgg taattattac tatttacaat
     5161 caaaggtggt cctgcaggaa gttcctatac tttctagaga ataggaactt cagatccact
     5221 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     5281 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttcgacc gaataaatac
     6421 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     6481 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     6541 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttgtcg agattttcag
     6601 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     6661 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     6721 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     6781 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattacgta
     6841 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     6901 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     6961 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     7021 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     7081 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     7141 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     7201 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     7261 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggttgct
     7321 acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc
     7381 ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg
     7441 tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc
     7501 tcaggctctc cccgtggagg taataattga cgatatgatc ctttttttct gatcaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     7921 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     7981 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     8041 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     8101 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     8161 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     8221 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     8281 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     8341 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     8401 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     8461 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     8521 atacgactca ctatagggcg aattg
//
