LOCUS       sequence-450702-        8343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the SCT1 allele from L-1374, under control of its
            native promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8343)
  AUTHORS   Paltenghi C, van Leeuwen J
  TITLE     Genetic suppression interactions are highly conserved across 
            genetically diverse yeast isolates.
  JOURNAL   G3 (Bethesda). 2025 Mar 3:jkaf047. doi: 10.1093/g3journal/jkaf047.
  PUBMED    40037589
REFERENCE   2  (bases 1 to 8343)
  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: "G3 
            (Bethesda). 2025 Mar 3:jkaf047. doi: 10.1093/g3journal/jkaf047."
COMMENT     Sequence Label: pRS313-SCT1-L1374
FEATURES             Location/Qualifiers
     source          1..8343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4535..4554)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4536..4554)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4561..4578)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4561..4577)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4570..4592)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4718..5173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4805..4824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5015..5036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5433..6095)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6096..6283)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6430..6449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6549..6571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6609..6627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6675..7178
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7208..7312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7313..7381,7382..8173)
                     /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(7531..7550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccccccta aaaagcccct gtgcgaggga
     1081 tatacataaa cattgagcat taatatttac aactcccaat gagcgataat gggtttatta
     1141 cataaataga acggcgaaag atttagtttc tagtatcttc accgtcagta aatggcatct
     1201 acgcatctcc ttctttccct tcttcttctt cttcctcttc ttcttcctct tcctcttccc
     1261 tggcagtatt ttcaccaatt ctcttgttta agacggcctg tgcgatttta cttgctaatc
     1321 catttttttc ctccaagatt tcgttttcta cctcaaattc ggaggaagaa gaaggtgcaa
     1381 cggatgtcga agctaaggaa gactctgact tacgatgaaa agtagaagag aataatggaa
     1441 tgttagagag ggaattgtca ctattgacca atgagacgcc atcggaatct tggttatggt
     1501 gttcataggc atcgtggttg tcacgttggc tgatgatagg tgatgacgaa tctttttctt
     1561 gtctttttat tttctgtttt cttagcattt tcctgcgatt caattctgag gtttttcgat
     1621 cttcttcctc ttcatcgatg acgtcgaatt cttcacggag ggcggcacta tcgaaatcgg
     1681 ggaataattc gcttccaaag ttatttacaa cttcgattat tctttctgcc agatttctac
     1741 gatccttttg tagcttttgc aagccctttg gagatgtaag agataaaacc agtggtctca
     1801 aagatttgaa accatccata ccaatatcac ccacgattaa ggctgaatat gtgactataa
     1861 cacacgagat gtaagacccg gaaaaaacat atattttatt ccatggttta tgtctgaggt
     1921 aataagtgat taaaacggac caaaagatgt aaagcaaggg cgcaaatccc atcccaatca
     1981 agattttcca cgtggcaatg acatcattag cctttatttt aactgtagac ttggacaaag
     2041 cggtacgggc cttttcttga gaaattctct tggctagaat gaagacaggt gagaacataa
     2101 tgatacctgg catggctaac gaaaagagga tgcagagccc gatggatcta aaaaaaacaa
     2161 gtccgaggtt ttttgcgaaa tttacctttg cctcctccac aaggtgatca ggaagattat
     2221 agtgtctcaa ggcggcatta tatgccatta tatctttggt caagtccgct attttaggat
     2281 cgtttctata gaattcgtaa cctttgacca ttcttctgtt catttccaca atcaagggca
     2341 acggtaactt ggtgctaaat tgtgtcatat atagtcttct tatcgtttga accaccatca
     2401 aagtttcata atcagaacat gtaacggtaa cggattgtaa acccttcgat atggtatcta
     2461 ataactcttt cactgcatct ctgttcgttt ccgggttgtg gtacttggcg actagttcct
     2521 tcggtatttc aatggggtca ccaaattcaa caaccgctct cgacctgaac ttatgtggat
     2581 ggaaataatt cataccacag ggaacaatct taacattgac gtcaggatgc ttatccatgc
     2641 aaccaagagc cataatcgcc acacctgctt tcaggggcaa caagtttgtt ctgtcgtggg
     2701 acccaccttc aggaaagatc ccaatgcagt tgttatgggc caaatgctca aaaactctat
     2761 ggtaaacgca agattggtcg actttggcgg catatttata agtagtgccg gtgagtaagg
     2821 cggttttaat ctctggtttg gccattttga actcttttct tagggtcaaa ctcgtgtcac
     2881 tttctatgga ctggatttct ccaaatccca ttgatttggg taacccgatg agaccctttg
     2941 gcatacaatc agtcaagaaa tgcgtgtcgt ggccgataac tctcttgtag tctgttggat
     3001 ctacgcggat agtaccttct gccggtttca aattatcctg cggccttacc acgcctatgg
     3061 ccatgaagaa actagccaaa aaccctatgg cgggttgctt taatgagctc tccgcaatca
     3121 agaaggacac acgtctgttg acagatttct tcacctcgcc cataaggatt acagggtcga
     3181 cgaactggtt agcatgcgga gccgcaacaa agatcacggg tccctgttgc ggtaccttga
     3241 aactgccccg gcctctgatt tctctaaaga agcagtggaa gatgttgtac aacagccatg
     3301 tggcgatgga gtaaagtatc ttcttggtag cgctaggctc ttggtagatg tatgcctgat
     3361 cagccgacgt cttgacgctt ttggcctcga tggaactgta attcgtagtt ttctgtgtgc
     3421 tcttggaaga ggcagatttc tccgtgagtt ttggtgcagg catttttttt taccccgatg
     3481 taaacgcccc tgtgctcttt ttactatata tttaattccg ggcttcctac acttaattca
     3541 ctctgttatg aactacaagc cattactaac atcaaccttt ccgaatcata aataaactgc
     3601 aactgggaaa gaaggcgggc ctacttttct ttttcttcat agcgcgtagg gcttggcagc
     3661 catagttgtt gagagaccga tggatgaggg attaaaagaa gggagaagaa gaatactaaa
     3721 agaggaatgt agagccgggt tacaggaaaa ctaacttttc actcagggcg ccgcatcttt
     3781 tccgcaggcg cgccatgaac tggccagcac gttctacggc gaaaccttcg aactggaggc
     3841 ggctcacaca tagtagtaag ttgccgcgac aggtgaccaa aatacaatct ttaatatcat
     3901 catatttgaa catgcccggt ttactgccat tatctagacg tgcgtctctc atcactttac
     3961 actcatgaaa caatattcgt tttaatactg aattttgagc gtcctttctt gccgctgtcg
     4021 cttctttgaa cgcgtgcaga ggaccgagcg tttcgagttt gttaagtagt tcggcggcag
     4081 aatcgcgcgc ccaatgaatg cgtttatctt ctgttgttat acggtgggcg taactgggtt
     4141 tgtaagctgt tggcgcttgc acacggttct gtggtagata taacctctca cgaagcgttt
     4201 ggcccaaaag ttgggcgcca agggcgccca actggtccat cagtatggca gtcctccgag
     4261 gaaggccctc agaattgaaa tctgccgtcg aatcattgac tctccctttt gacagcattg
     4321 ttgcgatcgc cagtggctcc gtctgcgcta caattgcacc atggtcaaac cgatccggat
     4381 gcagtgtctg tatagttaca ccggtgtaag tgtcaccctc cagcagagct cgctggatag
     4441 gtgcactgcc tttatgtctg ggaagtagcg aaggatggac gtttagcgcc cactagttct
     4501 agagcggccg ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc
     4561 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4621 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4681 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4741 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4801 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4861 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4921 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4981 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5041 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5101 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5161 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5221 gcatagatcc gtcgagttca agagaaaaaa aaagaaaaag caaaaagaaa aaaggaaagc
     5281 gcgcctcgtt cagaatgaca cgtatagaat gatgcattac cttgtcatct tcagtatcat
     5341 actgttcgta tacatactta ctgacattca taggtataca tatatacaca tgtatatata
     5401 tcgtatgctg cagctttaaa taatcggtgt cactacataa gaacaccttt ggtggaggga
     5461 acatcgttgg taccattggg cgaggtggct tctcttatgg caaccgcaag agccttgaac
     5521 gcactctcac tacggtgatg atcattcttg cctcgcagac aatcaacgtg gagggtaatt
     5581 ctgctagcct ctgcaaagct ttcaagaaaa tgcgggatca tctcgcaaga gagatctcct
     5641 actttctccc tttgcaaacc aagttcgaca actgcgtacg gcctgttcga aagatctacc
     5701 accgctctgg aaagtgcctc atccaaaggc gcaaatcctg atccaaacct ttttactcca
     5761 cgcacggccc ctagggcctc tttaaaagct tgaccgagag caatcccgca gtcttcagtg
     5821 gtgtgatggt cgtctatgtg taagtcacca atgcactcaa cgattagcga ccagccggaa
     5881 tgcttggcca gagcatgtat catatggtcc agaaacccta tacctgtgtg gacgttaatc
     5941 acttgcgatt gtgtggcctg ttctgctact gcttctgcct ctttttctgg gaagatcgag
     6001 tgctctatcg ctaggggacc accctttaaa gagatcgcaa tctgaatctt ggtttcattt
     6061 gtaatacgct ttactagggc tttctgctct gtcatctttg ccttcgttta tcttgcctgc
     6121 tcatttttta gtatattctt cgaagaaatc acattacttt atataatgta taattcatta
     6181 tgtgataatg ccaatcgcta agaaaaaaaa agagtcatcc gctaggtgga aaaaaaaaaa
     6241 tgaaaatcat taccgaggca taaaaaaata tagagtgtac tagaggaggc caagagtaat
     6301 agaaaaagaa aattgcggga aaggactgtg ttatgacttc cctgactaat gccgtgttca
     6361 aacgatacct ggcagtgact cctagcgctc accaagctct taaaacggga atttatggtg
     6421 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6481 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6541 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6601 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6661 ttagggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     6721 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     6781 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     6841 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     6901 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     6961 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     7021 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     7081 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     7141 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tcaggtggca cttttcgggg
     7201 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7261 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7501 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7621 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7861 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7921 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8161 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8341 ttc
//
