LOCUS       sequence-363309-        8942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E. coli PPK from yeast GPD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8942)
  TITLE     Downey Lab Plasmids
REFERENCE   2  (bases 1 to 8942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRS415-GPD-PPK
FEATURES             Location/Qualifiers
     source          1..8942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2800..3443
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3337..3360
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3449..3465
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3449..3465
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      5570..5817
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5586..5604)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5835..5854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5836..5854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5864..5881)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5864..5880)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5873..5895)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6021..6476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6108..6127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6318..6339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6777..7184
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7185..8279
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(8773..8792)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8892..8914
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcagtttat cattatcaat actcgccatt
     2821 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     2881 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     2941 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     3001 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     3061 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     3121 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     3181 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     3241 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     3301 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     3361 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     3421 aaaacaccag aacttagttt cgacggattc tagaactagt ggatcccccg ggctgcagga
     3481 attcgatatc aagcttatgg gtcaggaaaa gctatacatc gaaaaagagc tcagttggtt
     3541 atcgttcaat gaacgcgtgc ttcaggaagc ggcggacaaa tctaacccgc tgattgaaag
     3601 gatgcgtttc ctggggatct attccaataa ccttgatgag ttctataaag tccgcttcgc
     3661 tgaactgaag cgacgcatca ttattagcga agaacaaggc tccaactctc attcccgcca
     3721 tttactgggc aaaattcagt cccgggtgct gaaagccgat caggaattcg acggcctcta
     3781 caacgagcta ttgctggaga tggcgcgcaa ccagatcttc ctgattaatg aacgccagct
     3841 ctccgtcaat caacaaaact ggctgcgtca ttattttaag cagtatctgc gtcagcacat
     3901 tacgccgatt ttaatcaatc ctgacactga cttagtgcag ttcctgaaag atgattacac
     3961 ctatctggcg gtggaaatta tccgtggcga taccatccgt tacgcgctgc tggagatccc
     4021 atcagataaa gtgccgcgct ttgtgaattt accgccagaa gcgccgcgtc gacgcaagcc
     4081 gatgattctt ctggataaca ttctgcgtta ctgccttgat gatattttca aaggcttctt
     4141 tgattatgac gcgctgaatg cctattcaat gaagatgacc cgcgatgccg aatacgattt
     4201 agtgcatgag atggaagcca gcctgatgga gttgatgtct tccagtctca agcagcgttt
     4261 aactgctgag ccggtgcgtt ttgtttatca gcgcgatatg cccaatgcgc tggttgaagt
     4321 gttacgcgaa aaactgacta tttcccgcta cgactccatc gtccccggcg gtcgttatca
     4381 taattttaaa gactttatta atttccccaa tgtcggcaaa gccaatctgg tgaacaaacc
     4441 actgccgcgt ttacgccata tttggtttga taaagcccag ttccgcaatg gttttgatgc
     4501 cattcgcgaa cgcgatgtgt tgctctatta tccttatcac acctttgagc atgtgctgga
     4561 actgctgcgt caggcttcgt tcgacccgag cgtactggcg attaaaatta acatttaccg
     4621 cgtggcgaaa gattcacgca tcatcgactc gatgatccac gccgcacata acggtaagaa
     4681 agtcaccgtg gtggttgagt tacaggcgcg tttcgacgaa gaagccaaca ttcactgggc
     4741 gaagcgcctg accgaagcag gcgtgcacgt tatcttctct gcgccggggc tgaaaattca
     4801 cgccaaactg ttcctgattt cacgtaaaga aaacggtgaa gtggtgcgtt atgcacacat
     4861 cgggaccggg aactttaacg aaaaaaccgc gcgtctttat actgactatt cgttgctgac
     4921 cgccgatgcg cgcatcacca acgaagtacg gcgggtattt aactttattg aaaacccata
     4981 ccgtccggtg acatttgatt atttaatggt atcgccgcaa aactcccgcc gcctattgta
     5041 tgaaatggtg gaccgcgaga tcgccaacgc gcagcaaggg ctgcccagtg gtatcaccct
     5101 gaagctaaat aaccttgtcg ataaaggcct ggttgatcgt ctgtatgcgg cctccagctc
     5161 cggcgtaccg gttaatctgc tggttcgcgg aatgtgttcg ctgatcccca atctggaagg
     5221 cattagcgac aacattcgtg ccatcagtat tgttgaccgt taccttgaac atgaccgggt
     5281 ttatattttt gaaaatggcg gcgataaaaa ggtctacctt tcttccgccg actggatgac
     5341 gcgcaatatt gattatcgta ttgaagtggc gacgccgctg ctcgatccgc gcctgaagca
     5401 gcgggtactg gacatcatcg acatattgtt cagcgatacg gtcaaagcac gttatatcga
     5461 taaagaactc agtaatcgct acgttccccg cggcaatcgc cgcaaagtac gggcgcagtt
     5521 ggcgatttat gactacatca aatcactcga acaacctgaa taactcgagt catgtaatta
     5581 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     5641 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     5701 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     5761 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     5821 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     5881 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5941 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6001 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6181 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6241 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6301 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6361 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6421 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     6481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga
     6541 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     6601 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     6661 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     6721 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     6781 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     6841 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     6901 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     6961 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     7021 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     7081 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
     7141 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     7201 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     7261 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     7321 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     7381 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
     7441 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     7501 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     7561 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     7621 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     7681 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     7741 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     7801 tggaggaaac catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt
     7861 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     7921 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     7981 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     8041 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     8101 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     8161 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     8221 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     8281 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     8341 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     8401 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     8461 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     8521 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
     8581 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
     8641 atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg
     8701 tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgatatg
     8761 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     8821 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     8881 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     8941 ga
//
