LOCUS       p416-SHS-TIM                8328 bp    DNA     linear   SYN 30-MAY-2017
DEFINITION  LMBP 11196.
ACCESSION   MF048871
VERSION     MF048871.1
KEYWORDS    .
SOURCE      synthetic construct (code 12)
  ORGANISM  synthetic construct (code 12)
            other sequences; artificial sequences.
REFERENCE   1  (bases 1 to 8328)
  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  JOURNAL   Unpublished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 8328)
  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..8328
                     /organism="synthetic construct (code 12)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2005482"
                     /clone="p416-SHS-TIM"
     misc_feature    1..34
                     /note="FLP recombination target sequence FRT"
     regulatory      39..586
                     /regulatory_class="promoter"
                     /note="CaMAL2 promoter; derived from Candida albicans"
     CDS             join(593..858,1023..2028)
                     /note="FLP containing CaTUB2 intron"
                     /codon_start=1
                     /transl_table=12
                     /product="site-specific recombinase"
                     /protein_id="ART66091.1"
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWM
                     ITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEF
                     TIIPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEK
                     ILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCL
                     VTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLK
                     DNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKR
                     ASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKG
                     SAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      2029..2416
                     /regulatory_class="terminator"
                     /note="CaACT1 terminator; derived from Candida albicans"
     regulatory      2423..2920
                     /regulatory_class="promoter"
                     /note="CaACT1 promoter; derived from Candida albicans"
     gene            2921..3995
                     /gene="SAT1"
                     /note="5' region; derived from C. albicans codon-optimized
                     SAT1 gene"
     repeat_region   3682..3995
                     /gene="SAT1"
                     /rpt_type=direct
     gene            4002..6260
                     /gene="HygB"
                     /note="codon-optimized for C. albicans"
     regulatory      4002..4642
                     /regulatory_class="promoter"
                     /gene="HygB"
                     /note="CaTEF2 promoter; derived from C. albicans"
     CDS             4649..5674
                     /gene="HygB"
                     /codon_start=1
                     /transl_table=12
                     /product="hygromycin resistance marker"
                     /protein_id="ART66092.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
                     LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
                     EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
                     SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
                     TRPRAKE"
     regulatory      5681..6260
                     /regulatory_class="terminator"
                     /gene="HygB"
                     /note="CaPFK2 terminator; derived from C. albicans"
     gene            6261..6726
                     /gene="SAT1"
                     /note="3' region; derived from C. albicans codon-optimized
                     SAT1 gene"
     repeat_region   6261..6574
                     /gene="SAT1"
                     /rpt_type=direct
     misc_feature    6864..6897
                     /note="FLP recombination target sequence FRT"
     regulatory      6926..8325
                     /regulatory_class="promoter"
                     /note="CaMET3 promoter; derived from Candida albicans"
     misc_feature    8326..8328
                     /note="Start codon"
ORIGIN      
        1 gaagttccta tactttctag agaataggaa cttcggatcc aataatgatt ggtttgatat
       61 ttttgtctag taccatctgt accattacac ttaaattatc tttatatctg tctaactcga
      121 ctgtctggat ttcattgatg tagtcgtatg catcgttagt tccaaaaaat attgtcatca
      181 atttgatatt ggtttccgac tctaaaattt ttggaagaat ttgtctagcg tgctctgagt
      241 tgtagccact gaaaccacgg ttaataacat ccaattttcg gatatacaca ttctgtaatg
      301 ctggatgaaa gccatactgg gtacaactaa actgggtgat ggagtcaccg aacaacacaa
      361 atttaccgta ttccatgatt gctatggttg agaatttttt ttttttcttg tcccacgcca
      421 tttttcaaat tatgcagttg agaatgttag tttttgtgta caccccgttc gctgaatatt
      481 tcggaataat tcaaagattg gggagtgggg gaggcgatag acgaagacac ggtataaaaa
      541 tgggcaaaat tttccccaac tttttgcagt ggtttaacta ataatcgtcg acatgccaca
      601 atttgatata ttatgtaaaa caccacctaa ggtgcttgtt cgtcagtttg tggaaaggtt
      661 tgaaagacct tcaggtgaga aaatagcatt atgtgctgct gaactaacct atttatgttg
      721 gatgattaca cataacggaa cagcaatcaa gagagccaca ttcatgagct ataatactat
      781 cataagcaat tcgttgagtt tcgatattgt caataaatca ctccagttta aatacaagac
      841 gcaaaaagca acaattttgt aggtatagac actgaagaaa aaaaaattct atcattgtta
      901 tgttgtgatc tttgatcttt agttgtcggg ttaacacctg ccaattggat caatacatca
      961 atcaattaat tctaatcttg aaaaaaaaat tcattttact aacaattttt tcttttattt
     1021 agggaagcct cattaaagaa attgattcct gcttgggaat ttacaattat tccttactat
     1081 ggacaaaaac atcaatctga tatcactgat attgtaagta gtttgcaatt acagttcgaa
     1141 tcatcggaag aagcagataa gggaaatagc cacagtaaaa aaatgcttaa agcacttcta
     1201 agtgagggtg aaagcatctg ggagatcact gagaaaatac taaattcgtt tgagtatact
     1261 tcgagattta caaaaacaaa aactttatac caattcctct tcctagctac tttcatcaat
     1321 tgtggaagat tcagcgatat taagaacgtt gatccgaaat catttaaatt agtccaaaat
     1381 aagtatttgg gagtaataat ccagtgttta gtgacagaga caaagacaag cgttagtagg
     1441 cacatatact tctttagcgc aaggggtagg atcgatccac ttgtatattt ggatgaattt
     1501 ttgaggaatt ctgaaccagt cctaaaacga gtaaatagga ccggcaattc ttcaagcaat
     1561 aaacaggaat accaattatt aaaagataac ttagtcagat cgtacaataa agctttgaag
     1621 aaaaatgcgc cttattcaat ctttgctata aaaaatggcc caaaatctca cattggaaga
     1681 catttgatga cctcatttct ttcaatgaag ggcctaacgg agttgactaa tgttgtggga
     1741 aattggagcg ataagcgtgc ttctgccgtg gccaggacaa cgtatactca tcagataaca
     1801 gcaatacctg atcactactt cgcactagtt tctcggtact atgcatatga tccaatatca
     1861 aaggaaatga tagcattgaa ggatgagact aatccaattg aggagtggca gcatatagaa
     1921 cagctaaagg gtagtgctga aggaagcata cgataccccg catggaatgg gataatatca
     1981 caggaggtac tagactacct ttcatcctac ataaatagac gcatataaga gtgaaattct
     2041 ggaaatctgg aaatctggtt ttgtattctt gttattcttc tttttgttat tacatatata
     2101 acttgttact tttttaaaaa aatctttgta tattttataa atatataaaa ctaaatttaa
     2161 gaaaaagaga aaaatgtttt atttgagaga ttgaaatttt acttgaattt agcttagctt
     2221 ttataaagta ttattatgta aaaaaacaaa acaaatatac attaaaaagt taagactata
     2281 aaatagccac ccaaggcatt tctatatctt gttgttgttg ttttcatctt ctgtatcaga
     2341 ggaacttatt ttattatttt cgtcacgggt attttctctt gtttgatgat tcatcccatt
     2401 cattccatca taaaatgtcg agcgtcaaaa ctagagaata ataaagaaaa cgatcttttc
     2461 aaaaagaaaa aaccttttag ttttcctttg ttgttgttgt gggtgtgtgc tatttatatt
     2521 atatagttta ctcataatac cataaaatat tcggtttgat taggttattt taataagcta
     2581 atttgtttct aatcgtgtaa tttatgctgt gtatattaag tagtgtgtgc actgcccaaa
     2641 aatgtttgtt gtttatagtc ggttaaagag aaaaaagaaa aaaagatcca tacacacacg
     2701 ttaattagtt gttcaacgta atacactcat attttgttct tatttgcttt cggtcgctgt
     2761 tctcaccaag atttattgcc aacgaaacaa ttttttttta tatattttca gatttttctt
     2821 tttttccttt cctttccttt tctaattttc actcctggtt ttctttcttt cttagaaaca
     2881 ttatctcgat attaatatta aaaaaatata atcattcaaa atggacggtg gtatgtttta
     2941 gtttagcttc aattctaatt gattgattaa tcagttgatt ggtttcaata tgacaaatgg
     3001 gtagggtggg aaaacttcat tttcaattca gatcaaactt ttttgttgtc gacataatat
     3061 ttctcgtttg ggatgttact gtcacattaa taatacacac acatcagctt ataattttga
     3121 aagtaattta tcagatatgt tgtgacgatc aatggaaatg gctaacttca atgtatctgt
     3181 tcttcccctt tttcaaagtt cacgtttttt gattgattga ttgatctgtc ggcagtggtt
     3241 tcaaaaccat tcggtgagta atcctatcaa tcaatgttac gacaaaaggc tcaatattca
     3301 aaattgcaat gttttatgtt ttcctacgtg tacttgtgca aggcaattga ttcaacattg
     3361 cttttggtgt ttgacgagtt tctagtttgg acttgtgttg ttatctgggc tatacagatt
     3421 tcccggctca ctatgaattt tttttttcga cgctcagtgc acacaactat aaacaacaca
     3481 aacacaaaca cagcaagaaa aaaaaaaaac gaacattgaa ttgaaaccaa gccaactgaa
     3541 aaattcctta tttaaatgac tgtcatacta acccattttt atagaagaag ttgctgcttt
     3601 agttatcgat aacggttctc atatgaaaat ttcggtgatc cctgagcagg tggcggaaac
     3661 attggatgct gagaaccatt tcattgttcg tgaagtgttc gatgtgcacc tatccgacca
     3721 aggctttgaa ctatctacca gaagtgtgag cccctaccgg aaggattaca tctcggatga
     3781 tgactctgat gaagactctg cttgctatgg cgcattcatc gaccaagagc ttgtcgggaa
     3841 gattgaactc aactcaacat ggaacgatct agcctctatc gaacacattg ttgtgtcgca
     3901 cacgcaccga ggcaaaggag tcgcgcacag tctcatcgaa tttgcgaaaa agtgggcact
     3961 aagcagacag ctccttggca tacgattaga gacacgacgt cgtatagtgc ttgctgttcg
     4021 atattgctag aggcaaagaa aaaggattac ctcacgccta aaatggaatt tgaaaattga
     4081 ggagatgagt tgagcttttg tcttttacac tttccctatt caattgtgcc gcaaaaactt
     4141 caatatctaa cacagtgtga aatatttgtt atgtataccg aaagccaaat accatttaaa
     4201 ctggtatttc aagatttcat aaatcaagtt tctggttaaa ttaattgctg ttaaacagaa
     4261 caagagcttc acattgatgt tggaataact actatagaat tataaaatac acaaaaccag
     4321 gtcgtgtacg caataacttt tcaatgtatg aaaaaatttt tttttttttt ttgcttcttc
     4381 tcttcgcttc ttttttctag ttatcaattt ggcaatgaat tgatttgatt gccctttgtt
     4441 tagtttacta tgaaaaaaaa tttttttttt gtctctgtca gattttctct catttttttt
     4501 tttgcctcat aatgaacaac tcaaaaaaga atataaattt ctcatcaaag acctttccaa
     4561 ttgaaaattt tttctttcct tcttttcctt tttttttttc tttctttctt cttttctaag
     4621 tatacatttt atataatcaa tcctcgagat gaaaaaacca gaattgactg ctacttctgt
     4681 tgaaaaattt ttgattgaaa aatttgattc tgtttctgat ttgatgcaat tgtctgaagg
     4741 tgaagaatca agagcttttt cttttgatgt tggtggtaga ggttatgttt tgagagttaa
     4801 ttcttgtgct gatggttttt ataaagatag atatgtttat agacattttg cttctgctgc
     4861 tttgccaatt ccagaagttt tggatattgg tgaattttct gaatctttga cttattgtat
     4921 ttcaagaaga gctcaaggtg ttactttgca agatttgcca gaaactgaat tgccagctgt
     4981 tttgcaacca gttgctgaag ctatggatgc tattgctgct gctgatttgt ctcaaacttc
     5041 tggttttggt ccatttggtc cacaaggtat tggtcaatat actacttgga gagattttat
     5101 ttgtgctatt gctgatccac atgtttatca ttggcaaact gttatggatg atactgtttc
     5161 tgcttctgtt gctcaagcat tggatgaatt gatgttgtgg gctgaagatt gtccagaagt
     5221 tagacatttg gttcatgctg attttggttc taataatgtt ttgactgata atggtagaat
     5281 tactgctgtt attgattggt ctgaagctat gtttggtgat tctcaatatg aagttgctaa
     5341 tatttttttt tggagaccat ggttggcttg tatggaacaa caaactagat attttgaaag
     5401 aagacatcca gaattggctg gttctccaag attgagagct tatatgttga gaattggttt
     5461 ggatcaattg tatcaatctt tggttgatgg taattttgat gatgctgctt gggctcaagg
     5521 tagatgtgat gctattgtta gatcaggtgc tggtactgtt ggtagaactc aaattgctag
     5581 aagatcagct gctgtttgga ctgatggttg tgttgaagtt ttggctgatt ctggtaatag
     5641 aagaccatct actagaccaa gagctaaaga ataaggatcc gacgaagttg gtgtatgata
     5701 acgaatttat agtttttata ttgtaaaata atgaaatatt tgtttttttt ttactttgat
     5761 gtatatgctg tatcccaatc ctcgatacat tgcagaattg aatcatttgg tttatctgtt
     5821 gttttgtgct aagtaagata ttttccatct agaagtgata gctttttatt atcaaattac
     5881 aatccctatt acatgtggac atcaactcca tttctttgcg cattttatag tagagttaga
     5941 gttagcttgt cacatctcgt tgcttccttt tgctgcaact gtttccgatt tcaaaatgaa
     6001 aatagcttaa agttacccgg agggcctgca cataatcaaa atatagtaaa ttcccaaaag
     6061 aacagtcgtg tgcagtttcc aaagtgaaat tttttttttg tttctctcac ttccttgtaa
     6121 ggttgtttcc ctcttctttc tctctaacca atcctttgta gattgaaaat tttttcttgc
     6181 aaatttttct ttaactctct ttcccttttt ttttcgaatc aaggtaccat atcattacca
     6241 aattagacat catgcctatg cattgttcgt gaagtgttcg atgtgcacct atccgaccaa
     6301 ggctttgaac tatctaccag aagtgtgagc ccctaccgga aggattacat ctcggatgat
     6361 gactctgatg aagactctgc ttgctatggc gcattcatcg accaagagct tgtcgggaag
     6421 attgaactca actcaacatg gaacgatcta gcctctatcg aacacattgt tgtgtcgcac
     6481 acgcaccgag gcaaaggagt cgcgcacagt ctcatcgaat ttgcgaaaaa gtgggcacta
     6541 agcagacagc tccttggcat acgattagag acacaaacga acaatgtacc tgcctgcaat
     6601 ttgtacgcaa aatgtggctt tactctcggc ggcattgacc tcttcacgta taaaactaga
     6661 cctcaagtct cgaacgaaac agcgatgtac tggtactggt tctcgggagc acaggatgac
     6721 gcctaacata tgtgaagtgt gaagggggag attttcactt tattagattt gtatatatgt
     6781 ataataaata aataaataag ttaaataaat aattagataa gggtggtaat tattactatt
     6841 tacaatcaaa ggtggtcctg caggaagttc ctatactttc tagagaatag gaacttcaga
     6901 tccactagtt ctagagcggt ttaaacaatg actattggta gcgtgttcta tttttcttat
     6961 gcgctgaaaa actacgaaca attgtctatt ccaagcctgt ggcttttttt ggtgattcac
     7021 aatacataaa ttggcaaata cctcttctat agactttaac ctttgatgaa atggtatgga
     7081 taaaccactt ttgtggtcag ggtatatata tttttattcc aatctggcct ttataaattc
     7141 aaatttccaa ggggactctg ctaaaccttc tgatataact ataactctaa taatggctgt
     7201 aaaaagtgtg aaaaaaagat gatattcaca aaaaagataa aggggatttc atattgggta
     7261 tctagattta aacatcgtat ttcaacaatg aaactagaat caatacgaaa tccagggcac
     7321 aactgtgttc attcccttgg gaattatact gagtaatggg aagaatgtgc aaaaaatacc
     7381 cgtgtcttat ataatttcca tttgtgtagg gcttaaaagt ggacaacttc aggttacgaa
     7441 accacgaaaa tgtctaaacg tttgatcaaa ttcgtaactc atggaaaatc acagcaacaa
     7501 attaaggatg gatatatccg gtacttacaa ataaaaacac taaagaagta taaattttga
     7561 actgaaattc tatagaaaat cttttttttt tgggtaaaat tatgcgattg tggctcatag
     7621 taacgtaatc atacagttca acaaacatca ttttgtcaaa ggatttctaa tcctattctt
     7681 tttctacttg gcttttgagt atatatatat atacatcctt tgttcatcat catgttcgat
     7741 tgaatttttt tcttggttgt tattagattg tctacttgag tttttcttgg tgcaaattgt
     7801 ggtgtttttc taacaaattt ttcaacattt aaagtatttc tggaactttt ctacaaggaa
     7861 cgacatagat ccattaatgc gcccctctaa aacaataccc ataatgtctt cttaagtatg
     7921 catctgctta taaatgtttc tggatttact ttagttagaa tgtcgacacg acaatgaact
     7981 attgctgtag atcacgtgcg atcactgaat tatgagtttc tttttatttt tctttttggg
     8041 cttttttttt ttgcaatttt ttttttccac ctttgccaat tgcgtagtca catataatgt
     8101 atacaatttt tgcaactctg acactataat tgatccaata ataaagtata taatagatca
     8161 tcaagtatac gtaatctccc caaaaaaaaa attgcataaa ataaaatcta taaacctttt
     8221 ttagattttt agttgttcct tttttctttc tattcttcaa cttccaagtg ttgtcacttt
     8281 cttcctgtta actatattta aaagtaaata ccctccccag aaaacatg
//