LOCUS       62056_9160        8137 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM010, complete sequence.
ACCESSION   MK431395
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8137)
  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 8137)
  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 8137)
  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..8137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     22..55
                     /label=FLP recombination target sequence (FRT)
                     /note="FLP recombination target sequence (FRT)"
     misc_feature    62..1241
                     /label=promoter region from Candida albicans PCK1
                     /note="promoter region from Candida albicans PCK1"
     CDS             1242..2510
                     /label=FLP
                     /note="site-specific recombinase"
     misc_feature    2514..2901
                     /label=terminator region from Candida albicans ACT1
                     /note="terminator region from Candida albicans ACT1"
     misc_feature    2908..3405
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            3406..4632
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(3406..3415,4070..4632)
                     /codon_start=1
                     /transl_table=11
                     /gene="SAT1"
                     /product="streptomycin acetyltransferase"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="QBY25790.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     misc_feature    4636..4765
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(4770..4803)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(4805..4821)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4858..4876)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4897..4913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4921..4937)
                     /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(4945..4975)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4990..5011)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5299..5887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6107..6209
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6210..6866
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7357..7462)
                     /label=AmpR promoter
     rep_origin      7489..7944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8085..8101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8111..8129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggaagttcct atactttcta gagaatagga acttcggatc
       61 caatatatgc gagtcttttt tttccagtac ttttcttgca aaatctttac aaaaaaaaaa
      121 agaaatacaa aatgcaacca ccgagataag tatccccaac tgttacgtaa cacatcttac
      181 tactctgcaa cagccttggc caacactact actggtcgtt ggtcgttttc tggcaatgcc
      241 ccggaaaaac cttaaaactg ttatttggtc cattgccgtc aagcaaccaa aaataacaat
      301 ggcggtcccg gaagatgcag cgtaccaaga aaagagggga taaccgatgt gtttgtcacc
      361 actaagccat ctagcaagtc tatgagctca aatcaacaca gaagagaaaa tcaaatccac
      421 acatgcttgg tagcactagg cgaattagag ttttcagttt tcattcttat tgttaagtgt
      481 tgattgagca acataaaaag ccttgtccac tttctcgagg cagttaccgg aatattacgt
      541 attctcctcc ttctccccta ttttctaaac caacaaacaa aaaataaata tcatgagaag
      601 ttaaagaaaa taaatcgctc aaagtttatc tacgcaaaac tatgctaaaa caacaaacca
      661 caaaaaaaaa aaaaagaaat tgtcgacagc cgtaatagag tctagaccat ctgctcatta
      721 cccgaatgag ctggtcagta tctttcaaaa ttattggtta accaacaata ccgtcaactc
      781 aagtcctaac ctcttcttgc caacccctct agcaccccaa ggaacaatcg aatgtttctt
      841 ccttctctct tgaaacattc ttaataatac gtatgtaatt atttgctgac ttcttttttt
      901 tttgtttttt tcttattttt tagaacttcc gtttccccgc tcctactaaa atcttcccgg
      961 gttgaacggt aagtctcgga atacacttcc taataattcc tatacaccaa ttctacccca
     1021 gattttttgg aggttgcgcg taacctattc atatgccccc gggacaaggg aagcaatctt
     1081 aaaaagaaaa aaaattttcc agcaaaattc gttgatacaa gtctttatta atggtctgat
     1141 aaattagtat aaaaccacca tcaaattccc ttgaaaactt gattgatttt tttctttctt
     1201 tcttcgaatc aatcattaac ttattcaaat aattatcaat catgccacaa tttgatatat
     1261 tatgtaaaac accacctaag gtgcttgttc gtcagtttgt ggaaaggttt gaaagacctt
     1321 caggtgagaa aatagcatta tgtgctgctg aactaaccta tttatgttgg atgattacac
     1381 ataacggaac agcaatcaag agagccacat tcatgagcta taatactatc ataagcaatt
     1441 cgttgagttt cgatattgtc aataaatcac tccagtttaa atacaagacg caaaaagcaa
     1501 caattttgga agcctcatta aagaaattga ttcctgcttg ggaatttaca attattcctt
     1561 actatggaca aaaacatcaa tctgatatca ctgatattgt aagtagtttg caattacagt
     1621 tcgaatcatc ggaagaagca gataagggaa atagccacag taaaaaaatg cttaaagcac
     1681 ttctaagtga gggtgaaagc atctgggaga tcactgagaa aatactaaat tcgtttgagt
     1741 atacttcgag atttacaaaa acaaaaactt tataccaatt cctcttccta gctactttca
     1801 tcaattgtgg aagattcagc gatattaaga acgttgatcc gaaatcattt aaattagtcc
     1861 aaaataagta tttgggagta ataatccagt gtttagtgac agagacaaag acaagcgtta
     1921 gtaggcacat atacttcttt agcgcaaggg gtaggatcga tccacttgta tatttggatg
     1981 aatttttgag gaattctgaa ccagtcctaa aacgagtaaa taggaccggc aattcttcaa
     2041 gcaataaaca ggaataccaa ttattaaaag ataacttagt cagatcgtac aataaagctt
     2101 tgaagaaaaa tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa tctcacattg
     2161 gaagacattt gatgacctca tttctttcaa tgaagggcct aacggagttg actaatgttg
     2221 tgggaaattg gagcgataag cgtgcttctg ccgtggccag gacaacgtat actcatcaga
     2281 taacagcaat acctgatcac tacttcgcac tagtttctcg gtactatgca tatgatccaa
     2341 tatcaaagga aatgatagca ttgaaggatg agactaatcc aattgaggag tggcagcata
     2401 tagaacagct aaagggtagt gctgaaggaa gcatacgata ccccgcatgg aatgggataa
     2461 tatcacagga ggtactagac tacctttcat cctacataaa tagacgcata taagagtgaa
     2521 attctggaaa tctggaaatc tggttttgta ttcttgttat tcttcttttt gttattacat
     2581 atataacttg ttactttttt aaaaaaatct ttgtatattt tataaatata taaaactaaa
     2641 tttaagaaaa agagaaaaat gttttatttg agagattgaa attttacttg aatttagctt
     2701 agcttttata aagtattatt atgtaaaaaa acaaaacaaa tatacattaa aaagttaaga
     2761 ctataaaata gccacccaag gcatttctat atcttgttgt tgttgttttc atcttctgta
     2821 tcagaggaac ttattttatt attttcgtca cgggtatttt ctcttgtttg atgattcatc
     2881 ccattcattc catcataaaa tgtcgagcgt caaaactaga gaataataaa gaaaacgatc
     2941 ttttcaaaaa gaaaaaacct tttagttttc ctttgttgtt gttgtgggtg tgtgctattt
     3001 atattatata gtttactcat aataccataa aatattcggt ttgattaggt tattttaata
     3061 agctaatttg tttctaatcg tgtaatttat gctgtgtata ttaagtagtg tgtgcactgc
     3121 ccaaaaatgt ttgttgttta tagtcggtta aagagaaaaa agaaaaaaag atccatacac
     3181 acacgttaat tagttgttca acgtaataca ctcatatttt gttcttattt gctttcggtc
     3241 gctgttctca ccaagattta ttgccaacga aacaattttt ttttatatat tttcagattt
     3301 ttcttttttt cctttccttt ccttttctaa ttttcactcc tggttttctt tctttcttag
     3361 aaacattatc tcgatattaa tattaaaaaa atataatcat tcaaaatgga cggtggtatg
     3421 ttttagttta gcttcaattc taattgattg attaatcagt tgattggttt caatatgaca
     3481 aatgggtagg gtgggaaaac ttcattttca attcagatca aacttttttg ttgtcgacat
     3541 aatatttctc gtttgggatg ttactgtcac attaataata cacacacatc agcttataat
     3601 tttgaaagta atttatcaga tatgttgtga cgatcaatgg aaatggctaa cttcaatgta
     3661 tctgttcttc ccctttttca aagttcacgt tttttgattg attgattgat ctgtcggcag
     3721 tggtttcaaa accattcggt gagtaatcct atcaatcaat gttacgacaa aaggctcaat
     3781 attcaaaatt gcaatgtttt atgttttcct acgtgtactt gtgcaaggca attgattcaa
     3841 cattgctttt ggtgtttgac gagtttctag tttggacttg tgttgttatc tgggctatac
     3901 agatttcccg gctcactatg aatttttttt ttcgacgctc agtgcacaca actataaaca
     3961 acacaaacac aaacacagca agaaaaaaaa aaaacgaaca ttgaattgaa accaagccaa
     4021 ctgaaaaatt ccttatttaa atgactgtca tactaaccca tttttataga agaagttgct
     4081 gctttagtta tcgataacgg ttctcatatg aaaatttcgg tgatccctga gcaggtggcg
     4141 gaaacattgg atgctgagaa ccatttcatt gttcgtgaag tgttcgatgt gcacctatcc
     4201 gaccaaggct ttgaactatc taccagaagt gtgagcccct accggaagga ttacatctcg
     4261 gatgatgact ctgatgaaga ctctgcttgc tatggcgcat tcatcgacca agagcttgtc
     4321 gggaagattg aactcaactc aacatggaac gatctagcct ctatcgaaca cattgttgtg
     4381 tcgcacacgc accgaggcaa aggagtcgcg cacagtctca tcgaatttgc gaaaaagtgg
     4441 gcactaagca gacagctcct tggcatacga ttagagacac aaacgaacaa tgtacctgcc
     4501 tgcaatttgt acgcaaaatg tggctttact ctcggcggca ttgacctctt cacgtataaa
     4561 actagacctc aagtctcgaa cgaaacagcg atgtactggt actggttctc gggagcacag
     4621 gatgacgcct aacatatgtg aagtgtgaag ggggagattt tcactttatt agatttgtat
     4681 atatgtataa taaataaata aataagttaa ataaataatt agataagggt ggtaattatt
     4741 actatttaca atcaaaggtg gtcctgcagg aagttcctat actttctaga gaataggaac
     4801 ttcagatcca ctagttctag agcggccgcc accgcggtgg agctccagct tttgttccct
     4861 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5701 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttcga
     6001 ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg
     6061 ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcacgtaa
     6121 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttgt
     6181 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     6241 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     6301 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     6361 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     6421 cggaattacg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     6481 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     6541 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     6601 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
     6661 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     6721 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     6781 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     6841 actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa
     6901 cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag
     6961 caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
     7021 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
     7081 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tccttttttt
     7141 ctgatcaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7381 aagcatttat caagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     7501 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7561 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7621 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7681 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7741 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7801 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7861 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7921 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     7981 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8041 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     8101 tgagcgcgcg taatacgact cactataggg cgaattg
//
