LOCUS       p416-CtSHS                6847 bp    DNA     linear   SYN 30-MAY-2017
DEFINITION  LMBP 11194.
ACCESSION   MF048869
VERSION     MF048869.1
KEYWORDS    .
SOURCE      synthetic construct (code 12)
  ORGANISM  synthetic construct (code 12)
            other sequences; artificial sequences.
REFERENCE   1  (bases 1 to 6847)
  AUTHORS   Bijlani,S. and Ganesan,K.
  TITLE     A Cassette for One-Step Gene Deletion and Regulated Gene Expression
            in Clinical Isolates of Diploid Candida species
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6847)
  AUTHORS   Bijlani,S. and Ganesan,K.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2017) Yeast Molecular Biology, CSIR- Institute of
            Microbial Technology, Sector 39-A, Chandigarh, Chandigarh 160036,
            India
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6847
                     /organism="synthetic construct (code 12)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2005482"
                     /clone="p416-CtSHS"
     misc_feature    1..34
                     /note="FLP recombination target sequence FRT"
     regulatory      39..679
                     /regulatory_class="promoter"
                     /note="CtMAL2 promoter; derived from Candida tropicalis"
     gene            686..1957
                     /gene="FLP"
     CDS             686..1957
                     /gene="FLP"
                     /codon_start=1
                     /transl_table=12
                     /product="site-specific recombinase"
                     /protein_id="ART66087.1"
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWM
                     ITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEF
                     TIIPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEK
                     ILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCL
                     VTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLK
                     DNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKR
                     ASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKG
                     SAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      1958..2345
                     /regulatory_class="terminator"
                     /note="CaACT1 terminator; derived from Candida albicans"
     regulatory      2352..2849
                     /regulatory_class="promoter"
                     /note="CaACT1 promoter; derived from Candida albicans"
     gene            2850..3924
                     /gene="SAT1"
                     /note="5' region; derived from C. albicans codon-optimized
                     SAT1 gene"
     repeat_region   3611..3924
                     /gene="SAT1"
                     /rpt_type=direct
     gene            3931..6189
                     /gene="HygB"
                     /note="codon-optimized for C. albicans"
     regulatory      3931..4571
                     /regulatory_class="promoter"
                     /gene="HygB"
                     /note="CaTEF2 promoter; derived from C. albicans"
     CDS             4578..5603
                     /gene="HygB"
                     /note="hygromycin resistance marker"
                     /codon_start=1
                     /transl_table=12
                     /product="HygB"
                     /protein_id="ART66088.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
                     LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
                     EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
                     SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
                     TRPRAKE"
     regulatory      5610..6189
                     /regulatory_class="terminator"
                     /gene="HygB"
                     /note="CaPFK2 terminator; derived from C. albicans"
     gene            6190..6655
                     /gene="SAT1"
                     /note="3' region; derived from C. albicans codon-optimized
                     SAT1 gene"
     repeat_region   6190..6503
                     /gene="SAT1"
                     /rpt_type=direct
     misc_feature    6793..6826
                     /note="FLP recombination target sequence FRT"
ORIGIN      
        1 gaagttccta tactttctag agaataggaa cttcggatgg tagtttatag agaatataca
       61 aacaattggt gatttatatt tataaaagtt aaaagccctg tataccgaat tactaagaaa
      121 ttattgtctg atttgtaatt cagaacgcaa agtgtaatgc cctgtatctt attttttttt
      181 ttttgcagct atacgataag aatctcattt ttttggtgcc aatgcgatcg tttatacgca
      241 attcctaatt gaaacgtctc aacaaaaaaa ttgtaaatca tcttcaattg gtagctttaa
      301 taactctaaa ttcaaggggg ggattgccag cgtatctgta taaacgagtg ttcttttcta
      361 ctacattgct gtggatacta ttaatagaca aacagtagtg tgtgtataca catacagaat
      421 ccatctctct ccgccacatt gtttccaata aaaaaatatt atccagtaat tggtgttttt
      481 gtgtttctat ggatcaattc tcggaaaaat tatcgcgaga acaaagagca agtgtgggga
      541 gatggggagt caacacagca agagaacaaa agctttaaat agtccatctt ttcaacggac
      601 aagctgaaaa aaaagagcaa ttacttgtta ctcttcaatt ttttttttct ctacttttta
      661 tatattcttc aaacaaaacg tcgacatgcc acaatttgat atattatgta aaacaccacc
      721 taaggtgctt gttcgtcagt ttgtggaaag gtttgaaaga ccttcaggtg agaaaatagc
      781 attatgtgct gctgaactaa cctatttatg ttggatgatt acacataacg gaacagcaat
      841 caagagagcc acattcatga gctataatac tatcataagc aattcgttga gtttcgatat
      901 tgtcaataaa tcactccagt ttaaatacaa gacgcaaaaa gcaacaattt tggaagcctc
      961 attaaagaaa ttgattcctg cttgggaatt tacaattatt ccttactatg gacaaaaaca
     1021 tcaatctgat atcactgata ttgtaagtag tttgcaatta cagttcgaat catcggaaga
     1081 agcagataag ggaaatagcc acagtaaaaa aatgcttaaa gcacttctaa gtgagggtga
     1141 aagcatctgg gagatcactg agaaaatact aaattcgttt gagtatactt cgagatttac
     1201 aaaaacaaaa actttatacc aattcctctt cctagctact ttcatcaatt gtggaagatt
     1261 cagcgatatt aagaacgttg atccgaaatc atttaaatta gtccaaaata agtatttggg
     1321 agtaataatc cagtgtttag tgacagagac aaagacaagc gttagtaggc acatatactt
     1381 ctttagcgca aggggtagga tcgatccact tgtatatttg gatgaatttt tgaggaattc
     1441 tgaaccagtc ctaaaacgag taaataggac cggcaattct tcaagcaata aacaggaata
     1501 ccaattatta aaagataact tagtcagatc gtacaataaa gctttgaaga aaaatgcgcc
     1561 ttattcaatc tttgctataa aaaatggccc aaaatctcac attggaagac atttgatgac
     1621 ctcatttctt tcaatgaagg gcctaacgga gttgactaat gttgtgggaa attggagcga
     1681 taagcgtgct tctgccgtgg ccaggacaac gtatactcat cagataacag caatacctga
     1741 tcactacttc gcactagttt ctcggtacta tgcatatgat ccaatatcaa aggaaatgat
     1801 agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg
     1861 tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact
     1921 agactacctt tcatcctaca taaatagacg catataagag tgaaattctg gaaatctgga
     1981 aatctggttt tgtattcttg ttattcttct ttttgttatt acatatataa cttgttactt
     2041 ttttaaaaaa atctttgtat attttataaa tatataaaac taaatttaag aaaaagagaa
     2101 aaatgtttta tttgagagat tgaaatttta cttgaattta gcttagcttt tataaagtat
     2161 tattatgtaa aaaaacaaaa caaatataca ttaaaaagtt aagactataa aatagccacc
     2221 caaggcattt ctatatcttg ttgttgttgt tttcatcttc tgtatcagag gaacttattt
     2281 tattattttc gtcacgggta ttttctcttg tttgatgatt catcccattc attccatcat
     2341 aaaatgtcga gcgtcaaaac tagagaataa taaagaaaac gatcttttca aaaagaaaaa
     2401 accttttagt tttcctttgt tgttgttgtg ggtgtgtgct atttatatta tatagtttac
     2461 tcataatacc ataaaatatt cggtttgatt aggttatttt aataagctaa tttgtttcta
     2521 atcgtgtaat ttatgctgtg tatattaagt agtgtgtgca ctgcccaaaa atgtttgttg
     2581 tttatagtcg gttaaagaga aaaaagaaaa aaagatccat acacacacgt taattagttg
     2641 ttcaacgtaa tacactcata ttttgttctt atttgctttc ggtcgctgtt ctcaccaaga
     2701 tttattgcca acgaaacaat ttttttttat atattttcag atttttcttt ttttcctttc
     2761 ctttcctttt ctaattttca ctcctggttt tctttctttc ttagaaacat tatctcgata
     2821 ttaatattaa aaaaatataa tcattcaaaa tggacggtgg tatgttttag tttagcttca
     2881 attctaattg attgattaat cagttgattg gtttcaatat gacaaatggg tagggtggga
     2941 aaacttcatt ttcaattcag atcaaacttt tttgttgtcg acataatatt tctcgtttgg
     3001 gatgttactg tcacattaat aatacacaca catcagctta taattttgaa agtaatttat
     3061 cagatatgtt gtgacgatca atggaaatgg ctaacttcaa tgtatctgtt cttccccttt
     3121 ttcaaagttc acgttttttg attgattgat tgatctgtcg gcagtggttt caaaaccatt
     3181 cggtgagtaa tcctatcaat caatgttacg acaaaaggct caatattcaa aattgcaatg
     3241 ttttatgttt tcctacgtgt acttgtgcaa ggcaattgat tcaacattgc ttttggtgtt
     3301 tgacgagttt ctagtttgga cttgtgttgt tatctgggct atacagattt cccggctcac
     3361 tatgaatttt ttttttcgac gctcagtgca cacaactata aacaacacaa acacaaacac
     3421 agcaagaaaa aaaaaaaacg aacattgaat tgaaaccaag ccaactgaaa aattccttat
     3481 ttaaatgact gtcatactaa cccattttta tagaagaagt tgctgcttta gttatcgata
     3541 acggttctca tatgaaaatt tcggtgatcc ctgagcaggt ggcggaaaca ttggatgctg
     3601 agaaccattt cattgttcgt gaagtgttcg atgtgcacct atccgaccaa ggctttgaac
     3661 tatctaccag aagtgtgagc ccctaccgga aggattacat ctcggatgat gactctgatg
     3721 aagactctgc ttgctatggc gcattcatcg accaagagct tgtcgggaag attgaactca
     3781 actcaacatg gaacgatcta gcctctatcg aacacattgt tgtgtcgcac acgcaccgag
     3841 gcaaaggagt cgcgcacagt ctcatcgaat ttgcgaaaaa gtgggcacta agcagacagc
     3901 tccttggcat acgattagag acacgacgtc gtatagtgct tgctgttcga tattgctaga
     3961 ggcaaagaaa aaggattacc tcacgcctaa aatggaattt gaaaattgag gagatgagtt
     4021 gagcttttgt cttttacact ttccctattc aattgtgccg caaaaacttc aatatctaac
     4081 acagtgtgaa atatttgtta tgtataccga aagccaaata ccatttaaac tggtatttca
     4141 agatttcata aatcaagttt ctggttaaat taattgctgt taaacagaac aagagcttca
     4201 cattgatgtt ggaataacta ctatagaatt ataaaataca caaaaccagg tcgtgtacgc
     4261 aataactttt caatgtatga aaaaattttt tttttttttt tgcttcttct cttcgcttct
     4321 tttttctagt tatcaatttg gcaatgaatt gatttgattg ccctttgttt agtttactat
     4381 gaaaaaaaat tttttttttg tctctgtcag attttctctc attttttttt ttgcctcata
     4441 atgaacaact caaaaaagaa tataaatttc tcatcaaaga cctttccaat tgaaaatttt
     4501 ttctttcctt cttttccttt ttttttttct ttctttcttc ttttctaagt atacatttta
     4561 tataatcaat cctcgagatg aaaaaaccag aattgactgc tacttctgtt gaaaaatttt
     4621 tgattgaaaa atttgattct gtttctgatt tgatgcaatt gtctgaaggt gaagaatcaa
     4681 gagctttttc ttttgatgtt ggtggtagag gttatgtttt gagagttaat tcttgtgctg
     4741 atggttttta taaagataga tatgtttata gacattttgc ttctgctgct ttgccaattc
     4801 cagaagtttt ggatattggt gaattttctg aatctttgac ttattgtatt tcaagaagag
     4861 ctcaaggtgt tactttgcaa gatttgccag aaactgaatt gccagctgtt ttgcaaccag
     4921 ttgctgaagc tatggatgct attgctgctg ctgatttgtc tcaaacttct ggttttggtc
     4981 catttggtcc acaaggtatt ggtcaatata ctacttggag agattttatt tgtgctattg
     5041 ctgatccaca tgtttatcat tggcaaactg ttatggatga tactgtttct gcttctgttg
     5101 ctcaagcatt ggatgaattg atgttgtggg ctgaagattg tccagaagtt agacatttgg
     5161 ttcatgctga ttttggttct aataatgttt tgactgataa tggtagaatt actgctgtta
     5221 ttgattggtc tgaagctatg tttggtgatt ctcaatatga agttgctaat attttttttt
     5281 ggagaccatg gttggcttgt atggaacaac aaactagata ttttgaaaga agacatccag
     5341 aattggctgg ttctccaaga ttgagagctt atatgttgag aattggtttg gatcaattgt
     5401 atcaatcttt ggttgatggt aattttgatg atgctgcttg ggctcaaggt agatgtgatg
     5461 ctattgttag atcaggtgct ggtactgttg gtagaactca aattgctaga agatcagctg
     5521 ctgtttggac tgatggttgt gttgaagttt tggctgattc tggtaataga agaccatcta
     5581 ctagaccaag agctaaagaa taaggatccg acgaagttgg tgtatgataa cgaatttata
     5641 gtttttatat tgtaaaataa tgaaatattt gttttttttt tactttgatg tatatgctgt
     5701 atcccaatcc tcgatacatt gcagaattga atcatttggt ttatctgttg ttttgtgcta
     5761 agtaagatat tttccatcta gaagtgatag ctttttatta tcaaattaca atccctatta
     5821 catgtggaca tcaactccat ttctttgcgc attttatagt agagttagag ttagcttgtc
     5881 acatctcgtt gcttcctttt gctgcaactg tttccgattt caaaatgaaa atagcttaaa
     5941 gttacccgga gggcctgcac ataatcaaaa tatagtaaat tcccaaaaga acagtcgtgt
     6001 gcagtttcca aagtgaaatt ttttttttgt ttctctcact tccttgtaag gttgtttccc
     6061 tcttctttct ctctaaccaa tcctttgtag attgaaaatt ttttcttgca aatttttctt
     6121 taactctctt tccctttttt tttcgaatca aggtaccata tcattaccaa attagacatc
     6181 atgcctatgc attgttcgtg aagtgttcga tgtgcaccta tccgaccaag gctttgaact
     6241 atctaccaga agtgtgagcc cctaccggaa ggattacatc tcggatgatg actctgatga
     6301 agactctgct tgctatggcg cattcatcga ccaagagctt gtcgggaaga ttgaactcaa
     6361 ctcaacatgg aacgatctag cctctatcga acacattgtt gtgtcgcaca cgcaccgagg
     6421 caaaggagtc gcgcacagtc tcatcgaatt tgcgaaaaag tgggcactaa gcagacagct
     6481 ccttggcata cgattagaga cacaaacgaa caatgtacct gcctgcaatt tgtacgcaaa
     6541 atgtggcttt actctcggcg gcattgacct cttcacgtat aaaactagac ctcaagtctc
     6601 gaacgaaaca gcgatgtact ggtactggtt ctcgggagca caggatgacg cctaacatat
     6661 gtgaagtgtg aagggggaga ttttcacttt attagatttg tatatatgta taataaataa
     6721 ataaataagt taaataaata attagataag ggtggtaatt attactattt acaatcaaag
     6781 gtggtcctgc aggaagttcc tatactttct agagaatagg aacttcagat ccactagttc
     6841 tagagcg
//