LOCUS       40924_40592        5773 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSKPDR5-PGK1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  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   2  (bases 1 to 5773)
  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   3  (bases 1 to 5773)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5773)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "74"
COMMENT     SGRef: number: 2; 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: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5773
                     /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"
     terminator      1919..2189
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate
                     kinase gene"
     promoter        2225..2440
                     /label=URA3 promoter
     CDS             2441..3241
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    3265..3542
                     /label=3' end of Saccharomyces cerevisiae PDR5 ORF
                     /note="3' end of Saccharomyces cerevisiae PDR5 ORF"
     promoter        complement(3585..3603)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3624..3640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3648..3664)
                     /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(3672..3702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3717..3738)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4026..4614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4788..5645)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5646..5750)
                     /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 ataagctttc ccatgtctct actggtggtg gtgcttcttt
     1861 ggaattattg gaaggtaagg aattgccagg tgttgctttc ttatccgaaa agaaataaat
     1921 tgaattgaat tgaaatcgat agatcaattt ttttcttttc tctttcccca tcctttacgc
     1981 taaaataata gtttatttta ttttttgaat attttttatt tatatacgta tatatagact
     2041 attatttatc ttttaatgat tattaagatt tttattaaaa aaaaattcgc tcctctttta
     2101 atgcctttat gcagtttttt tttcccattc gatatttcta tgttcgggtt cagcgtattt
     2161 taagtttaat aactcgaaaa ttctgcgttc gttggatccg gtgattgatt gagcaagcta
     2221 gcttttcaat tcatcatttt ttttttattc ttttttttga tttcggtttc cttgaaattt
     2281 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     2341 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     2401 actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     2461 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     2521 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     2581 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     2641 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     2701 actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     2761 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     2821 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     2881 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     2941 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     3001 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     3061 tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg
     3121 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     3181 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     3241 ctaaaaaact gtattataag taaaaattca caccaccatc cggtatgaca tgtgggcagt
     3301 acatggaacc atatttacaa ctagcaaaga ctggttactt aactgatgaa aatgccactg
     3361 acacctgtag tttctgtcaa atatctacaa ccaatgatta cttagctaat gtcaattctt
     3421 tctacagtga gagatggaga aattatggta tcttcatctg ttatattgca ttcaattata
     3481 tcgctggtgt ctttttctac tggttagcaa gagtgcctaa aaagaacggt aaactctcca
     3541 agtctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     3601 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3661 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3721 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3781 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3841 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4441 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4621 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4681 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4741 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4801 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4861 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4921 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4981 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5041 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5101 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5161 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5221 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5281 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5341 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5401 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5461 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5521 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5581 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5641 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5701 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5761 cgaaaagtgc cac
//
