LOCUS       62056_9170        8738 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM019, complete sequence.
ACCESSION   MK431397
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8738)
  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 8738)
  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 8738)
  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..8738
                     /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..1842
                     /label=promoter region from Candida albicans PCK1
                     /note="promoter region from Candida albicans PCK1"
     CDS             1843..3111
                     /label=FLP
                     /note="site-specific recombinase"
     misc_feature    3115..3502
                     /label=terminator region from Candida albicans ACT1
                     /note="terminator region from Candida albicans ACT1"
     misc_feature    3509..4006
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            4007..5233
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(4007..4016,4671..5233)
                     /codon_start=1
                     /transl_table=11
                     /gene="SAT1"
                     /product="streptomycin acetyltransferase"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="QBY25794.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     misc_feature    5237..5366
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(5371..5404)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(5406..5422)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5459..5477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5498..5514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5522..5538)
                     /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(5546..5576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5591..5612)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5900..6488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6708..6810
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6811..7467
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7958..8063)
                     /label=AmpR promoter
     rep_origin      8090..8545
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8686..8702
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8712..8730
                     /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 ccaatatatg cgagtctttt ttttccagta cttttcttgc aaaatcttta caaaaaaaaa
      721 aagaaataca aaatgcaacc accgagataa gtatccccaa ctgttacgta acacatctta
      781 ctactctgca acagccttgg ccaacactac tactggtcgt tggtcgtttt ctggcaatgc
      841 cccggaaaaa ccttaaaact gttatttggt ccattgccgt caagcaacca aaaataacaa
      901 tggcggtccc ggaagatgca gcgtaccaag aaaagagggg ataaccgatg tgtttgtcac
      961 cactaagcca tctagcaagt ctatgagctc aaatcaacac agaagagaaa atcaaatcca
     1021 cacatgcttg gtagcactag gcgaattaga gttttcagtt ttcattctta ttgttaagtg
     1081 ttgattgagc aacataaaaa gccttgtcca ctttctcgag gcagttaccg gaatattacg
     1141 tattctcctc cttctcccct attttctaaa ccaacaaaca aaaaataaat atcatgagaa
     1201 gttaaagaaa ataaatcgct caaagtttat ctacgcaaaa ctatgctaaa acaacaaacc
     1261 acaaaaaaaa aaaaaagaaa ttgtcgacag ccgtaataga gtctagacca tctgctcatt
     1321 acccgaatga gctggtcagt atctttcaaa attattggtt aaccaacaat accgtcaact
     1381 caagtcctaa cctcttcttg ccaacccctc tagcacccca aggaacaatc gaatgtttct
     1441 tccttctctc ttgaaacatt cttaataata cgtatgtaat tatttgctga cttctttttt
     1501 ttttgttttt ttcttatttt ttagaacttc cgtttccccg ctcctactaa aatcttcccg
     1561 ggttgaacgg taagtctcgg aatacacttc ctaataattc ctatacacca attctacccc
     1621 agattttttg gaggttgcgc gtaacctatt catatgcccc cgggacaagg gaagcaatct
     1681 taaaaagaaa aaaaattttc cagcaaaatt cgttgataca agtctttatt aatggtctga
     1741 taaattagta taaaaccacc atcaaattcc cttgaaaact tgattgattt ttttctttct
     1801 ttcttcgaat caatcattaa cttattcaaa taattatcaa tcatgccaca atttgatata
     1861 ttatgtaaaa caccacctaa ggtgcttgtt cgtcagtttg tggaaaggtt tgaaagacct
     1921 tcaggtgaga aaatagcatt atgtgctgct gaactaacct atttatgttg gatgattaca
     1981 cataacggaa cagcaatcaa gagagccaca ttcatgagct ataatactat cataagcaat
     2041 tcgttgagtt tcgatattgt caataaatca ctccagttta aatacaagac gcaaaaagca
     2101 acaattttgg aagcctcatt aaagaaattg attcctgctt gggaatttac aattattcct
     2161 tactatggac aaaaacatca atctgatatc actgatattg taagtagttt gcaattacag
     2221 ttcgaatcat cggaagaagc agataaggga aatagccaca gtaaaaaaat gcttaaagca
     2281 cttctaagtg agggtgaaag catctgggag atcactgaga aaatactaaa ttcgtttgag
     2341 tatacttcga gatttacaaa aacaaaaact ttataccaat tcctcttcct agctactttc
     2401 atcaattgtg gaagattcag cgatattaag aacgttgatc cgaaatcatt taaattagtc
     2461 caaaataagt atttgggagt aataatccag tgtttagtga cagagacaaa gacaagcgtt
     2521 agtaggcaca tatacttctt tagcgcaagg ggtaggatcg atccacttgt atatttggat
     2581 gaatttttga ggaattctga accagtccta aaacgagtaa ataggaccgg caattcttca
     2641 agcaataaac aggaatacca attattaaaa gataacttag tcagatcgta caataaagct
     2701 ttgaagaaaa atgcgcctta ttcaatcttt gctataaaaa atggcccaaa atctcacatt
     2761 ggaagacatt tgatgacctc atttctttca atgaagggcc taacggagtt gactaatgtt
     2821 gtgggaaatt ggagcgataa gcgtgcttct gccgtggcca ggacaacgta tactcatcag
     2881 ataacagcaa tacctgatca ctacttcgca ctagtttctc ggtactatgc atatgatcca
     2941 atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga gtggcagcat
     3001 atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg gaatgggata
     3061 atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat ataagagtga
     3121 aattctggaa atctggaaat ctggttttgt attcttgtta ttcttctttt tgttattaca
     3181 tatataactt gttacttttt taaaaaaatc tttgtatatt ttataaatat ataaaactaa
     3241 atttaagaaa aagagaaaaa tgttttattt gagagattga aattttactt gaatttagct
     3301 tagcttttat aaagtattat tatgtaaaaa aacaaaacaa atatacatta aaaagttaag
     3361 actataaaat agccacccaa ggcatttcta tatcttgttg ttgttgtttt catcttctgt
     3421 atcagaggaa cttattttat tattttcgtc acgggtattt tctcttgttt gatgattcat
     3481 cccattcatt ccatcataaa atgtcgagcg tcaaaactag agaataataa agaaaacgat
     3541 cttttcaaaa agaaaaaacc ttttagtttt cctttgttgt tgttgtgggt gtgtgctatt
     3601 tatattatat agtttactca taataccata aaatattcgg tttgattagg ttattttaat
     3661 aagctaattt gtttctaatc gtgtaattta tgctgtgtat attaagtagt gtgtgcactg
     3721 cccaaaaatg tttgttgttt atagtcggtt aaagagaaaa aagaaaaaaa gatccataca
     3781 cacacgttaa ttagttgttc aacgtaatac actcatattt tgttcttatt tgctttcggt
     3841 cgctgttctc accaagattt attgccaacg aaacaatttt tttttatata ttttcagatt
     3901 tttctttttt tcctttcctt tccttttcta attttcactc ctggttttct ttctttctta
     3961 gaaacattat ctcgatatta atattaaaaa aatataatca ttcaaaatgg acggtggtat
     4021 gttttagttt agcttcaatt ctaattgatt gattaatcag ttgattggtt tcaatatgac
     4081 aaatgggtag ggtgggaaaa cttcattttc aattcagatc aaactttttt gttgtcgaca
     4141 taatatttct cgtttgggat gttactgtca cattaataat acacacacat cagcttataa
     4201 ttttgaaagt aatttatcag atatgttgtg acgatcaatg gaaatggcta acttcaatgt
     4261 atctgttctt cccctttttc aaagttcacg ttttttgatt gattgattga tctgtcggca
     4321 gtggtttcaa aaccattcgg tgagtaatcc tatcaatcaa tgttacgaca aaaggctcaa
     4381 tattcaaaat tgcaatgttt tatgttttcc tacgtgtact tgtgcaaggc aattgattca
     4441 acattgcttt tggtgtttga cgagtttcta gtttggactt gtgttgttat ctgggctata
     4501 cagatttccc ggctcactat gaattttttt tttcgacgct cagtgcacac aactataaac
     4561 aacacaaaca caaacacagc aagaaaaaaa aaaaacgaac attgaattga aaccaagcca
     4621 actgaaaaat tccttattta aatgactgtc atactaaccc atttttatag aagaagttgc
     4681 tgctttagtt atcgataacg gttctcatat gaaaatttcg gtgatccctg agcaggtggc
     4741 ggaaacattg gatgctgaga accatttcat tgttcgtgaa gtgttcgatg tgcacctatc
     4801 cgaccaaggc tttgaactat ctaccagaag tgtgagcccc taccggaagg attacatctc
     4861 ggatgatgac tctgatgaag actctgcttg ctatggcgca ttcatcgacc aagagcttgt
     4921 cgggaagatt gaactcaact caacatggaa cgatctagcc tctatcgaac acattgttgt
     4981 gtcgcacacg caccgaggca aaggagtcgc gcacagtctc atcgaatttg cgaaaaagtg
     5041 ggcactaagc agacagctcc ttggcatacg attagagaca caaacgaaca atgtacctgc
     5101 ctgcaatttg tacgcaaaat gtggctttac tctcggcggc attgacctct tcacgtataa
     5161 aactagacct caagtctcga acgaaacagc gatgtactgg tactggttct cgggagcaca
     5221 ggatgacgcc taacatatgt gaagtgtgaa gggggagatt ttcactttat tagatttgta
     5281 tatatgtata ataaataaat aaataagtta aataaataat tagataaggg tggtaattat
     5341 tactatttac aatcaaaggt ggtcctgcag gaagttccta tactttctag agaataggaa
     5401 cttcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     5461 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5521 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5581 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5641 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5701 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6301 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttcg
     6601 accgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct
     6661 gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta
     6721 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagttg
     6781 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     6841 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     6901 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     6961 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     7021 ccggaattac gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     7081 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     7141 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     7201 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     7261 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     7321 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     7381 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     7441 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
     7501 acgcctggtt gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa
     7561 gcaaattcga cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt
     7621 gctggtttac cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct
     7681 gaggccagtt tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcctttttt
     7741 tctgatcaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7981 gaagcattta tcaagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8041 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     8101 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8161 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8221 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8281 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     8341 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8401 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     8461 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     8521 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     8581 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8641 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8701 gtgagcgcgc gtaatacgac tcactatagg gcgaattg
//
