LOCUS       sequence-314523-        6778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overxpression of PDE1 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  TITLE     Yeast viruses
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6778)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGAL-PDE1
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      1591..1838
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1607..1625)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1856..1875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1857..1875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1885..1902)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1885..1901)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1894..1916)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2042..2497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2129..2148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2339..2360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            2614..3733
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        2614..2957
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2958..3530
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      3536..3733
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3812..3831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3991..4009)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4050..4553
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4590..4694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4695..4763,4764..5555)
                     /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(4913..4932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5726..6314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6215..6234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6468..6485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6602..6623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6638..6643,6644..6661,6662..6668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6676..6692
                     /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     6681..6703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6700..6716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6700..6716
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6735..6755
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6737..6755
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggattcta gaactagcat ggttgtattc
      481 gaaataacta tacttggggc caatggagga cccaccgaat acggaacaca gtgttttata
      541 cttaagcctg ctagaacaga ggatccagaa ttaatagctg tagacggtgg tgcaggaatg
      601 taccagctac gggagatgtt ggtccaaggg cgaaatgaaa atgaaggtga tgatgagctt
      661 gttccaagct tttatgaaca tgatcgagag ccaatagagt tttttattga ttccaaactg
      721 aacatacaaa agggattatc caagtccttg ctacaatcat taaagaggca gggagagcat
      781 tttgaaagcg ccaatacaat gaagaaagct tacgaggttt tccaaggaat taccgactac
      841 tacattaccc atccccatct ggaccatatt agtgggctgg tagtaaactc cccctcaata
      901 tatgagcagg aaaacagtaa aaaggaaact atttggggat taccgcatac tattgacgtt
      961 ttgcaaaagc atgtctttaa tgacttaata tggccagatt taacggcaga acgttcaaga
     1021 aaattgaagc taaaatgctt gaatccgaag gaagttcaaa agtgcactac ctttccttgg
     1081 gatgtaatac cgttcaaagt tcatcatggg ataggcgtca agactggcgc gccggtatac
     1141 agcaccttct atatcttcag agacaggaaa agcaaagact gtataatagt ttgcggagat
     1201 gttgagcagg accgcagaga atctgaagaa tctctacttg aagaattttg gtcttacgtt
     1261 gctgaaaata taccgcttgt gcatctcaag ggtatattag tcgaatgttc atgcccatta
     1321 tcttccaagc ccgagcaatt gtatggtcat ctatctccaa tatatttaat caacgaatta
     1381 tccagtttga acactttata caacagtagt aagggattaa gcggtttgaa tgttatcgtt
     1441 actcacgtta agtcaacacc tgctaaaaga gacccaagac taaccatact cgaagagtta
     1501 cgatttttag ctgaggagag gaacttagga gacttgagaa tatctattgc gctagaaggc
     1561 cacactttgt ttctataact cgacctcgag tcatgtaatt agttatgtca cgcttacatt
     1621 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1681 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     1741 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     1801 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     1861 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1921 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1981 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2041 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2101 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2161 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2221 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2281 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2341 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2401 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2461 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     2521 cgcatctgtg cggtatttca caccgcatag ggttaattaa ggcgcgccag atctgtttag
     2581 cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
     2641 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
     2701 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
     2761 aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg
     2821 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
     2881 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
     2941 tccgaacata aacaaccatg ggtaccactc ttgacgacac ggcttaccgg taccgcacca
     3001 gtgtcccggg ggacgccgag gccatcgagg cactggatgg gtccttcacc accgacaccg
     3061 tcttccgcgt caccgccacc ggggacggct tcaccctgcg ggaggtgccg gtggacccgc
     3121 ccctgaccaa ggtgttcccc gacgacgaat cggacgacga atcggacgac ggggaggacg
     3181 gcgacccgga ctcccggacg ttcgtcgcgt acggggacga cggcgacctg gcgggcttcg
     3241 tggtcgtctc gtactccggc tggaaccgcc ggctgaccgt cgaggacatc gaggtcgccc
     3301 cggagcaccg ggggcacggg gtcgggcgcg cgttgatggg gctcgcgacg gagttcgccc
     3361 gcgagcgggg cgccgggcac ctctggctgg aggtcaccaa cgtcaacgca ccggcgatcc
     3421 acgcgtaccg gcggatgggg ttcaccctct gcggcctgga caccgccctg tacgacggca
     3481 ccgcctcgga cggcgagcag gcgctctaca tgagcatgcc ctgcccctaa tcagtactga
     3541 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     3601 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     3661 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     3721 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtgtcga aaacgagctc
     3781 gaattcatcg atgattatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4021 atgtcatgat aataatggtt tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg
     4081 tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt
     4141 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa
     4201 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga
     4261 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac
     4321 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg
     4381 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac
     4441 atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat
     4501 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
     4561 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4681 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4741 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4801 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4861 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4921 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4981 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5041 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5101 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5161 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5221 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5281 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5341 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5401 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5461 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5521 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5581 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5641 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6361 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6421 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6481 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6541 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6601 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6661 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6721 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctctagta
//
