LOCUS       sequence-328012-        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"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      1003..1190
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1058..1082)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1220..1239)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1221..1239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1249..1266)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1249..1265)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1258..1280)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1406..1861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1493..1512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1703..1724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            1978..3334
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1978..2321
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2322..3131
                     /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     complement(2389..2408)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3137..3334
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3413..3432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3532..3554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3592..3610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3678..3782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3783..3851,3852..4643)
                     /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(4001..4020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4814..5402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5303..5322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5556..5573
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      181 tggcggccgc tgttgggtta agaatactgg gcaatttcat gtttcttcaa caccacatat
      241 gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct gctcggagat
      301 taccgaatca aaaaaatttc aaagaaaccg gaatcaaaaa aaagaacaaa aaaaaaaaag
      361 atgaattgaa aagctttatg gaccctgaaa ccacagccac attaaccttc tttgatggtc
      421 aaaacttatc cttcaccata aatatgcctc gcaaaaaagg taattaacat atatagaatt
      481 acattattta tgaaatatca tcactatctc ttagcatctt taatcctttt ctacatcaga
      541 taacttcggt ttgttatcat cgtctgtatt gtcatcaatt ggcgcagtag cgctgaagtc
      601 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
      661 gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc
      721 actgctgaca aacaaaagaa tggtatcaaa gctaacttca aaattagaca caacattgaa
      781 gatggtggtg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
      841 gtcttgttac cagacaacca ttacttatcc tatcaatctg ccttatccaa agatccaaac
      901 gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt
      961 attgatgaat tgtacaaata ggtacccgcc acttctaaat aagcgaattt cttatgattt
     1021 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1081 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     1141 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     1201 agggatggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     1261 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1321 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1381 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1441 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1501 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1561 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1621 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1681 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1741 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1801 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1861 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggttaa
     1921 ttaaggcgcg ccagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac
     1981 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
     2041 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
     2101 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
     2161 ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat
     2221 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
     2281 ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc
     2341 acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2401 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     2461 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2521 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2581 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg
     2641 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2701 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2761 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2821 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     2881 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     2941 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     3001 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     3061 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     3121 agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     3181 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     3241 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     3301 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     3361 tccagtgtcg aaaacgagct cgaattcatc gatgattatg gtgcactctc agtacaatct
     3421 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3481 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3781 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3901 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3961 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4021 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4081 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4201 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4321 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4381 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4621 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4801 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4861 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4921 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4981 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5041 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5101 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5161 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5221 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5281 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5341 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5401 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5461 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5521 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5581 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5641 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaat
//
