LOCUS       p416-SHS                6754 bp    DNA     linear   SYN 30-MAY-2017
DEFINITION  LMBP 11193.
ACCESSION   MF048868
VERSION     MF048868.1
KEYWORDS    .
SOURCE      synthetic construct (code 12)
  ORGANISM  synthetic construct (code 12)
            other sequences; artificial sequences.
REFERENCE   1  (bases 1 to 6754)
  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  JOURNAL   Unpublished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 6754)
  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..6754
                     /organism="synthetic construct (code 12)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2005482"
                     /clone="p416-SHS"
     misc_feature    1..34
                     /note="FLP recombination target sequence FRT"
     regulatory      39..586
                     /regulatory_class="promoter"
                     /note="CaMAL2 promoter; derived from Candida albicans"
     gene            593..1864
                     /gene="FLP"
     CDS             593..1864
                     /gene="FLP"
                     /codon_start=1
                     /transl_table=12
                     /product="site-specific recombinase"
                     /protein_id="ART66085.1"
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWM
                     ITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEF
                     TIIPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEK
                     ILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCL
                     VTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLK
                     DNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKR
                     ASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKG
                     SAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     regulatory      1865..2252
                     /regulatory_class="terminator"
                     /note="CaACT1 terminator; derived from Candida albicans"
     regulatory      2259..2756
                     /regulatory_class="promoter"
                     /note="CaACT1 promoter; derived from Candida albicans"
     gene            2757..3831
                     /gene="SAT1"
                     /note="5' region; derived from C, albicans codon-optimized
                     SAT1 gene"
     repeat_region   3518..3831
                     /gene="SAT1"
                     /rpt_type=direct
     gene            3838..6096
                     /gene="HygB"
                     /note="codon-optimized for C. albicans"
     regulatory      3838..4478
                     /regulatory_class="promoter"
                     /gene="HygB"
                     /note="CaTEF2 promoter; derived from C. albicans"
     CDS             4485..5510
                     /gene="HygB"
                     /codon_start=1
                     /transl_table=12
                     /product="hygromycin resistance marker"
                     /protein_id="ART66086.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
                     LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
                     EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
                     SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
                     TRPRAKE"
     regulatory      5517..6096
                     /regulatory_class="terminator"
                     /gene="HygB"
                     /note="CaPFK2 terminator; derived from C. albicans"
     gene            6097..6562
                     /gene="SAT1"
                     /note="3' region; derived from C. albicans codon-optimized
                     SAT1 gene"
     repeat_region   6097..6410
                     /gene="SAT1"
                     /rpt_type=direct
     misc_feature    6700..6733
                     /note="FLP recombination target sequence FRT"
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 acaattttgg aagcctcatt aaagaaattg attcctgctt gggaatttac
      901 aattattcct tactatggac aaaaacatca atctgatatc actgatattg taagtagttt
      961 gcaattacag ttcgaatcat cggaagaagc agataaggga aatagccaca gtaaaaaaat
     1021 gcttaaagca cttctaagtg agggtgaaag catctgggag atcactgaga aaatactaaa
     1081 ttcgtttgag tatacttcga gatttacaaa aacaaaaact ttataccaat tcctcttcct
     1141 agctactttc atcaattgtg gaagattcag cgatattaag aacgttgatc cgaaatcatt
     1201 taaattagtc caaaataagt atttgggagt aataatccag tgtttagtga cagagacaaa
     1261 gacaagcgtt agtaggcaca tatacttctt tagcgcaagg ggtaggatcg atccacttgt
     1321 atatttggat gaatttttga ggaattctga accagtccta aaacgagtaa ataggaccgg
     1381 caattcttca agcaataaac aggaatacca attattaaaa gataacttag tcagatcgta
     1441 caataaagct ttgaagaaaa atgcgcctta ttcaatcttt gctataaaaa atggcccaaa
     1501 atctcacatt ggaagacatt tgatgacctc atttctttca atgaagggcc taacggagtt
     1561 gactaatgtt gtgggaaatt ggagcgataa gcgtgcttct gccgtggcca ggacaacgta
     1621 tactcatcag ataacagcaa tacctgatca ctacttcgca ctagtttctc ggtactatgc
     1681 atatgatcca atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga
     1741 gtggcagcat atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg
     1801 gaatgggata atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat
     1861 ataagagtga aattctggaa atctggaaat ctggttttgt attcttgtta ttcttctttt
     1921 tgttattaca tatataactt gttacttttt taaaaaaatc tttgtatatt ttataaatat
     1981 ataaaactaa atttaagaaa aagagaaaaa tgttttattt gagagattga aattttactt
     2041 gaatttagct tagcttttat aaagtattat tatgtaaaaa aacaaaacaa atatacatta
     2101 aaaagttaag actataaaat agccacccaa ggcatttcta tatcttgttg ttgttgtttt
     2161 catcttctgt atcagaggaa cttattttat tattttcgtc acgggtattt tctcttgttt
     2221 gatgattcat cccattcatt ccatcataaa atgtcgagcg tcaaaactag agaataataa
     2281 agaaaacgat cttttcaaaa agaaaaaacc ttttagtttt cctttgttgt tgttgtgggt
     2341 gtgtgctatt tatattatat agtttactca taataccata aaatattcgg tttgattagg
     2401 ttattttaat aagctaattt gtttctaatc gtgtaattta tgctgtgtat attaagtagt
     2461 gtgtgcactg cccaaaaatg tttgttgttt atagtcggtt aaagagaaaa aagaaaaaaa
     2521 gatccataca cacacgttaa ttagttgttc aacgtaatac actcatattt tgttcttatt
     2581 tgctttcggt cgctgttctc accaagattt attgccaacg aaacaatttt tttttatata
     2641 ttttcagatt tttctttttt tcctttcctt tccttttcta attttcactc ctggttttct
     2701 ttctttctta gaaacattat ctcgatatta atattaaaaa aatataatca ttcaaaatgg
     2761 acggtggtat gttttagttt agcttcaatt ctaattgatt gattaatcag ttgattggtt
     2821 tcaatatgac aaatgggtag ggtgggaaaa cttcattttc aattcagatc aaactttttt
     2881 gttgtcgaca taatatttct cgtttgggat gttactgtca cattaataat acacacacat
     2941 cagcttataa ttttgaaagt aatttatcag atatgttgtg acgatcaatg gaaatggcta
     3001 acttcaatgt atctgttctt cccctttttc aaagttcacg ttttttgatt gattgattga
     3061 tctgtcggca gtggtttcaa aaccattcgg tgagtaatcc tatcaatcaa tgttacgaca
     3121 aaaggctcaa tattcaaaat tgcaatgttt tatgttttcc tacgtgtact tgtgcaaggc
     3181 aattgattca acattgcttt tggtgtttga cgagtttcta gtttggactt gtgttgttat
     3241 ctgggctata cagatttccc ggctcactat gaattttttt tttcgacgct cagtgcacac
     3301 aactataaac aacacaaaca caaacacagc aagaaaaaaa aaaaacgaac attgaattga
     3361 aaccaagcca actgaaaaat tccttattta aatgactgtc atactaaccc atttttatag
     3421 aagaagttgc tgctttagtt atcgataacg gttctcatat gaaaatttcg gtgatccctg
     3481 agcaggtggc ggaaacattg gatgctgaga accatttcat tgttcgtgaa gtgttcgatg
     3541 tgcacctatc cgaccaaggc tttgaactat ctaccagaag tgtgagcccc taccggaagg
     3601 attacatctc ggatgatgac tctgatgaag actctgcttg ctatggcgca ttcatcgacc
     3661 aagagcttgt cgggaagatt gaactcaact caacatggaa cgatctagcc tctatcgaac
     3721 acattgttgt gtcgcacacg caccgaggca aaggagtcgc gcacagtctc atcgaatttg
     3781 cgaaaaagtg ggcactaagc agacagctcc ttggcatacg attagagaca cgacgtcgta
     3841 tagtgcttgc tgttcgatat tgctagaggc aaagaaaaag gattacctca cgcctaaaat
     3901 ggaatttgaa aattgaggag atgagttgag cttttgtctt ttacactttc cctattcaat
     3961 tgtgccgcaa aaacttcaat atctaacaca gtgtgaaata tttgttatgt ataccgaaag
     4021 ccaaatacca tttaaactgg tatttcaaga tttcataaat caagtttctg gttaaattaa
     4081 ttgctgttaa acagaacaag agcttcacat tgatgttgga ataactacta tagaattata
     4141 aaatacacaa aaccaggtcg tgtacgcaat aacttttcaa tgtatgaaaa aatttttttt
     4201 ttttttttgc ttcttctctt cgcttctttt ttctagttat caatttggca atgaattgat
     4261 ttgattgccc tttgtttagt ttactatgaa aaaaaatttt ttttttgtct ctgtcagatt
     4321 ttctctcatt tttttttttg cctcataatg aacaactcaa aaaagaatat aaatttctca
     4381 tcaaagacct ttccaattga aaattttttc tttccttctt ttcctttttt tttttctttc
     4441 tttcttcttt tctaagtata cattttatat aatcaatcct cgagatgaaa aaaccagaat
     4501 tgactgctac ttctgttgaa aaatttttga ttgaaaaatt tgattctgtt tctgatttga
     4561 tgcaattgtc tgaaggtgaa gaatcaagag ctttttcttt tgatgttggt ggtagaggtt
     4621 atgttttgag agttaattct tgtgctgatg gtttttataa agatagatat gtttatagac
     4681 attttgcttc tgctgctttg ccaattccag aagttttgga tattggtgaa ttttctgaat
     4741 ctttgactta ttgtatttca agaagagctc aaggtgttac tttgcaagat ttgccagaaa
     4801 ctgaattgcc agctgttttg caaccagttg ctgaagctat ggatgctatt gctgctgctg
     4861 atttgtctca aacttctggt tttggtccat ttggtccaca aggtattggt caatatacta
     4921 cttggagaga ttttatttgt gctattgctg atccacatgt ttatcattgg caaactgtta
     4981 tggatgatac tgtttctgct tctgttgctc aagcattgga tgaattgatg ttgtgggctg
     5041 aagattgtcc agaagttaga catttggttc atgctgattt tggttctaat aatgttttga
     5101 ctgataatgg tagaattact gctgttattg attggtctga agctatgttt ggtgattctc
     5161 aatatgaagt tgctaatatt tttttttgga gaccatggtt ggcttgtatg gaacaacaaa
     5221 ctagatattt tgaaagaaga catccagaat tggctggttc tccaagattg agagcttata
     5281 tgttgagaat tggtttggat caattgtatc aatctttggt tgatggtaat tttgatgatg
     5341 ctgcttgggc tcaaggtaga tgtgatgcta ttgttagatc aggtgctggt actgttggta
     5401 gaactcaaat tgctagaaga tcagctgctg tttggactga tggttgtgtt gaagttttgg
     5461 ctgattctgg taatagaaga ccatctacta gaccaagagc taaagaataa ggatccgacg
     5521 aagttggtgt atgataacga atttatagtt tttatattgt aaaataatga aatatttgtt
     5581 ttttttttac tttgatgtat atgctgtatc ccaatcctcg atacattgca gaattgaatc
     5641 atttggttta tctgttgttt tgtgctaagt aagatatttt ccatctagaa gtgatagctt
     5701 tttattatca aattacaatc cctattacat gtggacatca actccatttc tttgcgcatt
     5761 ttatagtaga gttagagtta gcttgtcaca tctcgttgct tccttttgct gcaactgttt
     5821 ccgatttcaa aatgaaaata gcttaaagtt acccggaggg cctgcacata atcaaaatat
     5881 agtaaattcc caaaagaaca gtcgtgtgca gtttccaaag tgaaattttt tttttgtttc
     5941 tctcacttcc ttgtaaggtt gtttccctct tctttctctc taaccaatcc tttgtagatt
     6001 gaaaattttt tcttgcaaat ttttctttaa ctctctttcc cttttttttt cgaatcaagg
     6061 taccatatca ttaccaaatt agacatcatg cctatgcatt gttcgtgaag tgttcgatgt
     6121 gcacctatcc gaccaaggct ttgaactatc taccagaagt gtgagcccct accggaagga
     6181 ttacatctcg gatgatgact ctgatgaaga ctctgcttgc tatggcgcat tcatcgacca
     6241 agagcttgtc gggaagattg aactcaactc aacatggaac gatctagcct ctatcgaaca
     6301 cattgttgtg tcgcacacgc accgaggcaa aggagtcgcg cacagtctca tcgaatttgc
     6361 gaaaaagtgg gcactaagca gacagctcct tggcatacga ttagagacac aaacgaacaa
     6421 tgtacctgcc tgcaatttgt acgcaaaatg tggctttact ctcggcggca ttgacctctt
     6481 cacgtataaa actagacctc aagtctcgaa cgaaacagcg atgtactggt actggttctc
     6541 gggagcacag gatgacgcct aacatatgtg aagtgtgaag ggggagattt tcactttatt
     6601 agatttgtat atatgtataa taaataaata aataagttaa ataaataatt agataagggt
     6661 ggtaattatt actatttaca atcaaaggtg gtcctgcagg aagttcctat actttctaga
     6721 gaataggaac ttcagatcca ctagttctag agcg
//