LOCUS       40924_40597        5654 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSKPDR5-PPUS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Nakamura K, Niimi M, Niimi K, Holmes AR, Yates JE, Decottignies A, 
            Monk BC, Goffeau A, Cannon RD.
  TITLE     Functional expression of Candida albicans drug efflux pump Cdr1p in 
            a Saccharomyces cerevisiae strain deficient in membrane transporters
  JOURNAL   Antimicrob. Agents Chemother. 45 (12), 3366-3374 (2001)
  PUBMED    11709310
REFERENCE   2  (bases 1 to 5654)
  AUTHORS   Lamping E, Niimi M, Cannon RD.
  TITLE     Small, synthetic, GC-rich mRNA stem-loop modules 5' proximal to the 
            AUG start-codon predictably tune gene expression in yeast
  JOURNAL   Microb. Cell Fact. 12 (1), 74 (2013)
  PUBMED    23895661
REFERENCE   3  (bases 1 to 5654)
  AUTHORS   Lamping E, Niimi M, Cannon RD.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2011) Sir John Walsh Research Institute, 
            University of Otago, 310 Great King Street, Dunedin, Otago 9054, New
            Zealand
REFERENCE   4  (bases 1 to 5654)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5654)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
            Agents Chemother."; date: "2001"; volume: "45"; issue: "12"; pages: 
            "3366-3374"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "74"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (15-AUG-2011) Sir John Walsh Research Institute, University of 
            Otago, 310 Great King Street, Dunedin, Otago 9054, New Zealand"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     667..683
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1823..1864
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      1870..2069
                     /label=3' end of Saccharomyces cerevisiae PDR5 ORF
                     /note="3' end of Saccharomyces cerevisiae PDR5 ORF"
                     /regulatory_class="terminator"
     promoter        2106..2321
                     /label=URA3 promoter
     CDS             2322..3122
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     misc_feature    3146..3423
                     /label=3' end of Saccharomyces cerevisiae PDR5 ORF
                     /note="3' end of Saccharomyces cerevisiae PDR5 ORF"
     promoter        complement(3466..3484)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3505..3521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3529..3545)
                     /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)."
     promoter        complement(3553..3583)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3598..3619)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3907..4495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4669..5526)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5527..5631)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      661 ccccctcgag gtcgacggta tcgatggtcc gtcatatacg gttctccgtt gggcgctacc
      721 acacacatat atataagcct gaatgcagta gacaaatttc agatatcata caaaggaagc
      781 gttctctggg acccctttct atgtttagat ggttcactga acttgttcgt taacgtaaat
      841 atgtcttcct ctttgattcc aacaatgata ttataaggaa attatcgtca caatctaatc
      901 aacaaggaca aagaaaagaa aaagttgaga gagagaacaa aagttcaaat caaagaaaaa
      961 aaagaacatc aaattaccta ttacaataaa caattaagcc atactcgcaa caatttgcca
     1021 taatagaaag cagcacctcg ttggcgcagt cccttacata gtacacaaca tttatcactt
     1081 cacacaatca ggagtggaac tcaatggaaa acaacaccac acgtatgatc ttactaataa
     1141 aagaacatga acgttcctca gcgcgaacgt tcgcattctg cgccttcgag cacaggataa
     1201 gttgcaggaa gccatcacat ctatgcaacg attatcacga cacaaccttg ccgccgagaa
     1261 aacgtccgtg gagaaccatt cggtcgattg cttcccacgg aacgagtgga ctgaaactta
     1321 agactgcccc tctctttccg cggaatcgct catgccgcgg tgccacaaca ttttcagatt
     1381 tactaagact ccggtgagtg tgggctcacc cgcgggtcgt gatcacgatt cagcaccctt
     1441 tggactcgtg attccgtgga aaggtcagat ctgtattcct acttatggta atgtgctaaa
     1501 aaaagagaaa tgtctccgcg gaactcttct acgccgtggt acgatatctg ttgaacgtaa
     1561 tctgagcaat acaaacaagg cctctcctat acatatataa ttgtgatgtg cataacctta
     1621 tggctgttcg cttttattac cataccttag aatgaaatcc aaaagaaaaa agtcacgcaa
     1681 agttgcaaac atataacaac tgtgttagtt atcactcgac tttgttattc taattataaa
     1741 taaattggca actaggaact ttcgaaaaag aaattaaaga cccttttaag ttttcgtatc
     1801 cgctcgttcg aaagaccccg ataagcttga tatcgaattc ctgcagcccg ggggatccac
     1861 tagttctagg ggtctgcttg tcatttcatt caaccaagtt gcagaaagtg cagctaactt
     1921 agcctctttg ttgtttacaa tgtctttgtc tttttgtggt gttatgacta ccccaagtgc
     1981 catgcctaga ttttggatat tcatgtacag ggtttcacct ttgacttatt tcattcaggc
     2041 tctgttggct gttggtgttg ctaacgtagt cgacggatcc ggtgattgat tgagcaagct
     2101 agcttttcaa ttcatcattt tttttttatt cttttttttg atttcggttt ccttgaaatt
     2161 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2221 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     2281 aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa gctacatata
     2341 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     2401 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     2461 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     2521 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     2581 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     2641 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     2701 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     2761 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     2821 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     2881 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     2941 atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag
     3001 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3061 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3121 actaaaaaac tgtattataa gtaaaaattc acaccaccat ccggtatgac atgtgggcag
     3181 tacatggaac catatttaca actagcaaag actggttact taactgatga aaatgccact
     3241 gacacctgta gtttctgtca aatatctaca accaatgatt acttagctaa tgtcaattct
     3301 ttctacagtg agagatggag aaattatggt atcttcatct gttatattgc attcaattat
     3361 atcgctggtg tctttttcta ctggttagca agagtgccta aaaagaacgg taaactctcc
     3421 aagtctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3481 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4321 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5641 ccgaaaagtg ccac
//
