LOCUS       62056_9180        5412 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM025, complete sequence.
ACCESSION   MK431399
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  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 5412)
  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 5412)
  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..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    369..866
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            867..2093
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(867..876,1531..2093)
                     /codon_start=1
                     /transl_table=11
                     /gene="SAT1"
                     /product="streptomycin acetyltransferase"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="QBY25796.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     misc_feature    2097..2226
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     promoter        complement(2300..2318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2325..2341)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2479..2778
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             3130..3921
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4131..4502
                     /label=BleoR
                     /note="antibiotic-binding protein"
     rep_origin      4643..5231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gccctttcat cccattcatt ccatcataaa
      361 atgtcgagcg tcaaaactag agaataataa agaaaacgat cttttcaaaa agaaaaaacc
      421 ttttagtttt cctttgttgt tgttgtgggt gtgtgctatt tatattatat agtttactca
      481 taataccata aaatattcgg tttgattagg ttattttaat aagctaattt gtttctaatc
      541 gtgtaattta tgctgtgtat attaagtagt gtgtgcactg cccaaaaatg tttgttgttt
      601 atagtcggtt aaagagaaaa aagaaaaaaa gatccataca cacacgttaa ttagttgttc
      661 aacgtaatac actcatattt tgttcttatt tgctttcggt cgctgttctc accaagattt
      721 attgccaacg aaacaatttt tttttatata ttttcagatt tttctttttt tcctttcctt
      781 tccttttcta attttcactc ctggttttct ttctttctta gaaacattat ctcgatatta
      841 atattaaaaa aatataatca ttcaaaatgg acggtggtat gttttagttt agcttcaatt
      901 ctaattgatt gattaatcag ttgattggtt tcaatatgac aaatgggtag ggtgggaaaa
      961 cttcattttc aattcagatc aaactttttt gttgtcgaca taatatttct cgtttgggat
     1021 gttactgtca cattaataat acacacacat cagcttataa ttttgaaagt aatttatcag
     1081 atatgttgtg acgatcaatg gaaatggcta acttcaatgt atctgttctt cccctttttc
     1141 aaagttcacg ttttttgatt gattgattga tctgtcggca gtggtttcaa aaccattcgg
     1201 tgagtaatcc tatcaatcaa tgttacgaca aaaggctcaa tattcaaaat tgcaatgttt
     1261 tatgttttcc tacgtgtact tgtgcaaggc aattgattca acattgcttt tggtgtttga
     1321 cgagtttcta gtttggactt gtgttgttat ctgggctata cagatttccc ggctcactat
     1381 gaattttttt tttcgacgct cagtgcacac aactataaac aacacaaaca caaacacagc
     1441 aagaaaaaaa aaaaacgaac attgaattga aaccaagcca actgaaaaat tccttattta
     1501 aatgactgtc atactaaccc atttttatag aagaagttgc tgctttagtt atcgataacg
     1561 gttctcatat gaaaatttcg gtgatccctg agcaggtggc ggaaacattg gatgctgaga
     1621 accatttcat tgttcgtgaa gtgttcgatg tgcacctatc cgaccaaggc tttgaactat
     1681 ctaccagaag tgtgagcccc taccggaagg attacatctc ggatgatgac tctgatgaag
     1741 actctgcttg ctatggcgca ttcatcgacc aagagcttgt cgggaagatt gaactcaact
     1801 caacatggaa cgatctagcc tctatcgaac acattgttgt gtcgcacacg caccgaggca
     1861 aaggagtcgc gcacagtctc atcgaatttg cgaaaaagtg ggcactaagc agacagctcc
     1921 ttggcatacg attagagaca caaacgaaca atgtacctgc ctgcaatttg tacgcaaaat
     1981 gtggctttac tctcggcggc attgacctct tcacgtataa aactagacct caagtctcga
     2041 acgaaacagc gatgtactgg tactggttct cgggagcaca ggatgacgcc taacatatgt
     2101 gaagtgtgaa gggggagatt ttcactttat tagatttgta tatatgtata ataaataaat
     2161 aaataagtta aataaataat tagataaggg tggtaattat tactatttac aatcaaaggt
     2221 ggtcctgcag agggcgaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca
     2281 tctagagggc ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     2341 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2401 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2461 cgcagcctat acgtacggca gtttaaggtt tacacctata aaagagagag ccgttatcgt
     2521 ctgtttgtgg atgtacagag tgatattatt gacacgccgg ggcgacggat ggtgatcccc
     2581 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
     2641 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
     2701 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
     2761 ctgatgttct ggggaatata aatgtcaggc atgagattat caaaaaggat cttcacctag
     2821 atccttttca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     2881 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     2941 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     3001 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     3061 tcgccgccaa ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga
     3121 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3301 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3901 cgccttcttg acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt
     3961 ctccttacgc atctgtgcgg tatttcacac cgcatacagg tggcactttt cggggaaatg
     4021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat agcacgtgag gagggccacc atggccaagt
     4141 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
     4201 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
     4261 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
     4321 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
     4381 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
     4441 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
     4501 actgacacgt gctaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctgggcttt tgctggcctt
     5281 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5341 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5401 aggaagcgga ag
//
