LOCUS       sequence-320198-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for replacement of PEST sequence in strains with 
            pr-yEVenus-PEST-ADH1t construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  AUTHORS   Gligorovski V, Sadeghi A, Rahi SJ
  TITLE     Multidimensional characterization of inducible promoters and a 
            highly light-sensitive LOV-transcription factor.
  JOURNAL   Nat Commun. 2023 Jun 27;14(1):3810. doi: 10.1038/s41467-023-38959-8.
  PUBMED    37369667
REFERENCE   2  (bases 1 to 5689)
  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: "Nat 
            Commun."; date: "2023-06-27"; volume: "14(1):3810. doi"; pages: " 
            10.1038/s41467-023-38959-8"
COMMENT     Sequence Label: pVG97
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            complement(249..1605)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(249..446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(452..1261)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1175..1194
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1262..1605)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1722..2177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2071..2090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2303..2325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2317..2334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2318..2334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2344..2363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2344..2362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2393..2580)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2501..2525
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3429..3449)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3429..3447)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3468..3484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3468..3484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3481..3503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3492..3508
                     /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)."
     promoter        complement(join(3516..3522,3523..3540,3541..3546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3561..3582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3699..3716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3870..4458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3950..3969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4629..5420,5421..5489))
                     /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     5252..5271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5490..5594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5662..5680
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataatca tcgatgaatt cgagctcgtt ttcgacactg gatggcggcg ttagtatcga
      241 atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
      301 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
      361 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
      421 aaaacaagaa tctttttatt gtcagtactg attagaaaaa ctcatcgagc atcaaatgaa
      481 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
      541 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
      601 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
      661 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
      721 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
      781 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
      841 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
      901 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttgc
      961 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1021 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1081 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1141 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1201 aatcagcatc catgttggaa tttaatcgcg gcctcgaaac gtgagtcttt tccttaccca
     1261 tggttgttta tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag
     1321 tatatgaaag aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc
     1381 ggcgtgggga caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag
     1441 cgaggagccg taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga
     1501 ttatacatgg ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct
     1561 gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc
     1621 cggcggggac gaggcaagct aaacagatct ggcgcgcctt aattaaccct atgcggtgtg
     1681 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     1741 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1801 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1861 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1921 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1981 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2041 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2101 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2161 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2221 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2281 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     2341 gcgtaatacg actcactata gggcgaattg ggtaccatcc ctagcggatc tgccggtaga
     2401 ggtgtggtca ataagagcga cctcatacta tacctgagaa agcaacctga cctacaggaa
     2461 agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt aaaatttgta
     2521 tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata agaaattcgc
     2581 ttatttagaa gtggcgggta cctatttgta caattcatca ataccatggg taataccagc
     2641 agcagtaaca aattctaaca agaccatgtg gtctctcttt tcgtttggat ctttggataa
     2701 ggcagattga taggataagt aatggttgtc tggtaacaag actggaccat caccaattgg
     2761 agtattttgt tgataatggt cagctaattg aacaccacca tcttcaatgt tgtgtctaat
     2821 tttgaagtta gctttgatac cattcttttg tttgtcagca gtgatgtaaa cattgtgaga
     2881 gttatagttg tattccaatt tgtgacctaa aatgttacca tcttctttaa aatcaatacc
     2941 ttttaattcg attctattaa ctaaggtatc accttcaaac ttgacttcag cgctactgcg
     3001 ccaattgatg acaatacaga cgatgataac aaaccgaagt tatctgatgt agaaaaggat
     3061 taaagatgct aagagatagt gatgatattt cataaataat gtaattctat atatgttaat
     3121 tacctttttt gcgaggcata tttatggtga aggataagtt ttgaccatca aagaaggtta
     3181 atgtggctgt ggtttcaggg tccataaagc ttttcaattc atcttttttt tttttgttct
     3241 tttttttgat tccggtttct ttgaaatttt tttgattcgg taatctccga gcagaaggaa
     3301 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtgg tgttgaagaa
     3361 acatgaaatt gcccagtatt cttaacccaa cagcggccgc caccgcggtg gagctccagc
     3421 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     3481 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5641 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
