LOCUS       40924_19111        8618 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Functional analysis vector pFA-HAH2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8618)
  AUTHORS   Sharma S, Alfatah M, Bari VK, Rawal Y, Paul S, Ganesan K.
  TITLE     Sphingolipid Biosynthetic Pathway Genes FEN1 and SUR4 Modulate 
            Amphotericin B Resistance
  JOURNAL   Antimicrob. Agents Chemother. 58 (4), 2409-2414 (2014)
  PUBMED    24395234
REFERENCE   2  (bases 1 to 8618)
  AUTHORS   Paul S, Rawal Y, Alfatah M, Bari VK, Sharma S, Choudhury S, Malik S,
            Ganesan K.
  TITLE     Single-Transformation Gene Function Test by Conditional Expression 
            of Both Alleles of Candida albicans Genes
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 8618)
  AUTHORS   Paul S, Rawal Y, Alfatah M, Bari VK, Sharma S, Choudhury S, Malik S,
            Ganesan K.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JUL-2013) Institute of Microbial Technology, Sector 
            39-A, Chandigarh, UT 160036, India
REFERENCE   4  (bases 1 to 8618)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8618)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
            Agents Chemother."; date: "2014"; volume: "58"; issue: "4"; pages: 
            "2409-2414"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (03-JUL-2013) Institute of Microbial Technology, Sector 39-A, 
            Chandigarh, UT 160036, India"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..55
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            60..928
                     /gene="HIS1"
                     /label=HIS1
                     /note="5' region; derived from Candida albicans HIS1"
     repeat_region   575..928
                     /gene="HIS1"
     gene            934..2884
                     /gene="ARG4"
                     /label=ARG4
                     /note="derived from Candida albicans ARG4"
     CDS             1391..2797
                     /codon_start=1
                     /gene="ARG4"
                     /product="arg4"
                     /label=ARG4
                     /note="argininosuccinate lyase"
                     /protein_id="AGY46360.1"
                     /translation="MSQQQDKQPSENKLWGGRFTGATDPLMDLYNASLPYDKVMYDADL
                     TGTKVYTQGLNKLGLITTEELNLIHQGLEQIRQEWHDNKFIIKAGDEDIHTANERRLGE
                     IIGKNISGKVHTGRSRNDQVATDMRIFVRESLLNLSKILHQFITAILERAHKEIDVLMP
                     GYTHLQKAQPIRWAHWLSSYATYFTEDYKRLQEIITRVNQSPLGSGALAGHPYGIDREF
                     LAKGLGFDGVIGNSLTAVSDRDFVVESLFWSTLFMNHISRFSEDLIIYSSGEFGFIKLA
                     DAYSTGSSLMPQKKNPDSLELLRGKSGRVFGQLSGFLMSIKSIPSTYNKDMQEDKEPLF
                     DALTTVEHSILIATGVISTLLIDKQNMEKALTMDMLATDLADYLVRKGVPFRETHHISG
                     ECVRKAEEEKLSGIDQLSFEQFQQIDSRFEKDVMETFDFEASVERRDALGGTAKSAVLK
                     QLENLKSILS"
     gene            2889..3738
                     /gene="HIS1"
                     /label=HIS1
                     /note="3' region; derived from Candida albicans HIS1"
     repeat_region   2889..3242
                     /gene="HIS1"
     protein_bind    3739..3772
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(3831..3849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3859..3875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4020..4475
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4609..5409)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5410..5630)
                     /label=URA3 promoter
     misc_feature    complement(5895..6398)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        6435..6539
                     /label=AmpR promoter
     CDS             6540..7397
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7571..8159
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8447..8468
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8483..8513
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8521..8537
                     /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     8545..8561
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8582..8600
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cttcgtacgc tgcaggtcga ctaccgttcg tataatgtat gctatacgaa gttatggatc
       61 ctggaggatg aggagacaga agttagtagt aacaatttgg ggacagaaga ggatacagtg
      121 acatcagagg aagaagacag cgaaggtata gtcaaaaagc aataggacag caagacaatt
      181 tatttattta atagtttttt ataaatgtaa aatctatcac aacatgtgta cttaattttc
      241 tgtggtaggg gaggccattt cgttgaaacg tgtcgcgagc ggttgtgaga attgactcat
      301 tttgattatt acgcaaccat tgaaattttt tatttttttt tggtgaagat ttttcccaca
      361 caacttcttc ttttacttaa ccaccaacta ccgataatgg atttagtcaa tcatttacca
      421 gaccgtttgt tatttgctgt tcctaaaaag ggcagattat acgaaaaatg ctgtaactta
      481 ttgagtggtg ccgatataca gtttagaaga tctaatagat tagatatagc actttctaca
      541 aacttgccaa ttgcattaat cttcttgcct gcagctgata tcccagtttt cgttggagaa
      601 ggtaattgtg acttgggtat aactgggtta gaccaaatca aagaagctga acaattcgac
      661 aacatcgagg acttgttgga tttgaaattt ggttcatgca aattgcagat ccaagttcca
      721 gcagatggcg agtacgaaaa gccagaacag cttgttggaa agaaaattgt gtcttcattt
      781 acaaaattga gtaccgacta tttcaaacaa ttgtcagaca aacctactaa tatcagatat
      841 gtcggtggtt ccgttgaggc ttcttgtgcc ttgggtgttg ctgatgctat tgtcgatttg
      901 gttgaaagtg gtgaaactat gaaagcagga tccccccttt agtaagattt ttcaagagta
      961 gtctcaaata aacagaatga aggattgaag gttttattca ataaataatt ggacagaaag
     1021 ttattgcaca gttttaacaa caacaatttc aaaatttcaa tttatgacat attgaccgac
     1081 ataattcagt catttctaat attcaaataa caagacacct tgtataaatg aatccacaag
     1141 cgtatatgaa caaccaaatt ttatgactaa ctgtgggtcg tgtacttttg acgattgagt
     1201 catttttacc aattaggtaa ttgaatttta ctaatttgtc attaaccgcg ctggccatcc
     1261 caacaacaac aacaacaatc aatttaaaaa cttgacaagg caaaaaaaaa aaatttcatc
     1321 tttgaaaaat ttttcttcct tttcttcttc tgttctttac ataacagcta tcaagaataa
     1381 tcaattaata atgtcacaac aacaagataa acaacctagt gaaaataaat tatggggtgg
     1441 tcgtttcacc ggtgccactg atccattgat ggatttgtat aatgcttcat taccttatga
     1501 taaagttatg tatgatgctg atttaactgg tactaaagta tacacacaag gattaaacaa
     1561 attaggatta attaccactg aagaattgaa tttgattcat caaggattag aacaaatcag
     1621 acaagaatgg catgataata aattcattat taaggctggt gatgaggata ttcatactgc
     1681 taatgaacgt cgattaggag aaatcattgg gaaaaacatt tctggtaaag ttcacactgg
     1741 tagatcaaga aatgatcaag ttgctaccga tatgagaatt ttcgttcgtg aatcattact
     1801 taatttaagt aaaattttac atcaattcat cacggcaatt cttgaacgag cacataaaga
     1861 aatcgatgta ttaatgcctg gttatactca tttacaaaaa gctcaaccta ttagatgggc
     1921 ccattggtta agttcatatg caacttattt caccgaagat tataaacgtt tacaagaaat
     1981 tataactcgt gttaaccaat ctccattagg atctggtgct ttagcaggtc atccttatgg
     2041 tattgatcgt gaatttttag ctaaaggatt aggatttgat ggtgtcattg ggaattcttt
     2101 aactgcagtt tcagatcgtg attttgttgt tgaaagttta ttttggagta cattatttat
     2161 gaatcatatt tctcgattct cagaagatct aattatttat tctagtggtg aatttgggtt
     2221 cataaaatta gctgatgctt attctactgg ttcttcattg atgccacaaa agaaaaaccc
     2281 tgactcatta gaattattac gtgggaaaag tggtagagta tttggtcagt tatctggatt
     2341 tttaatgtcg atcaaatcta ttccatctac ttataataaa gatatgcaag aggataaaga
     2401 accattattt gatgcattga ctacagtaga acattcaatt ttaattgcta ctggtgtgat
     2461 ttctacttta ctgattgata aacaaaatat ggaaaaagct ttaactatgg atatgttggc
     2521 tactgattta gctgattatt tagttagaaa aggagttcca tttagagaaa ctcatcatat
     2581 atctggtgaa tgtgttagaa aagctgaaga agaaaaatta tctggaattg atcaattatc
     2641 ttttgaacaa tttcaacaaa ttgattctcg ttttgaaaaa gacgttatgg aaacttttga
     2701 ttttgaagct agtgtggaaa gaagagatgc tcttggtggt actgctaaaa gtgccgtttt
     2761 aaaacaatta gaaaatttaa aatcaatctt aagttaattt ataaatagtc atataataat
     2821 cacagtattg tttaacttac ataaaatata atcaattaac acagagatac cttgtactac
     2881 aatagtttct gatatcccag ttttcgttgg agaaggtaat tgtgacttgg gtataactgg
     2941 gttagaccaa atcaaagaag ctgaacaatt cgacaacatc gaggacttgt tggatttgaa
     3001 atttggttca tgcaaattgc agatccaagt tccagcagat ggcgagtacg aaaagccaga
     3061 acagcttgtt ggaaagaaaa ttgtgtcttc atttacaaaa ttgagtaccg actatttcaa
     3121 acaattgtca gacaaaccta ctaatatcag atatgtcggt ggttccgttg aggcttcttg
     3181 tgccttgggt gttgctgatg ctattgtcga tttggttgaa agtggtgaaa ctatgaaagc
     3241 agctgggtta aaggccatag aaaccatatt ggagacttcg gctcatttga tttcgtcaaa
     3301 aaaaagtaaa ttcccagaaa tggtcaatat aatcgtccaa agacttcaag gtgttttagc
     3361 tgcccaggaa tatgtcttgt gtaactacaa tgccccaaaa tccatccaag caaaatgttt
     3421 aaccattact ccaggcagaa gggccgctac tgtctctact ttggacaaac acagcgacga
     3481 cgaagaggac tgggttgcca tctcatccat ggttaataga aaggaaatcg gtaatgtaat
     3541 ggacgaattg aagaaagctg gtgcaaccga tatattggta ctcgagatat caaattgtag
     3601 agtttaaaag aagtgatagt ttctcataaa tattatacat atatacttat agatctactt
     3661 caatttgact caatctgtct tataccatct ttatctacca ccttgatgta cacaccttta
     3721 aagtcaatag gcattctcat aacttcgtat aatgtatgct atacgaacgg tagtttaaac
     3781 gagctcgaat tcatcgatga tatcagatcc actagtggta cccaattcgc cctatagtga
     3841 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3901 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga
     4021 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4081 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4141 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4201 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4261 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4321 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4381 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4441 cgcgaatttt aacaaaatat taacgtttac aatttcctga tgcggtattt tctccttacg
     4501 catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct
     4561 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
     4621 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
     4681 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
     4741 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
     4801 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
     4861 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
     4921 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
     4981 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
     5041 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
     5101 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     5161 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
     5221 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     5281 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     5341 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     5401 tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa
     5461 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     5521 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaaag
     5581 aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt
     5641 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5701 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5761 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5821 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5881 ggtttcttag gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg
     5941 gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag
     6001 aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt
     6061 aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt
     6121 aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt
     6181 ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca
     6241 agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa
     6301 aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt
     6361 aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt
     6421 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6481 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6541 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6601 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6661 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6721 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6781 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6841 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6901 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6961 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7021 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7081 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7141 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7201 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7261 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7321 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7381 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7681 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8281 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8341 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     8401 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttacc
     8461 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcctatg ttgtgtggaa
     8521 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     8581 caattaaccc tcactaaagg gaacaaaagc tggagctc
//
