LOCUS       sequence-450356-        8354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the SCT1 allele from NCYC110, under control of 
            its native promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8354)
  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 8354)
  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-NCYC110
FEATURES             Location/Qualifiers
     source          1..8354
                     /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(4546..4565)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4547..4565)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4572..4589)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4572..4588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4581..4603)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4729..5184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4816..4835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5026..5047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5444..6106)
                     /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(6107..6294)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6441..6460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6560..6582
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6620..6638)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6686..7189
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7219..7323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7324..7392,7393..8184)
                     /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(7542..7561)
                     /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 tggtaccggg ccccccctaa aaagcccctg tgcgagggat
     1081 atacataaac attgagcatt aatatttaca actcccaatg agcgataatg ggtttattac
     1141 ataaatagaa cggcgaaaga tttagtttct agtatcttca ccgtcagtaa atggcatcta
     1201 cgcatctcct tctttccctt cttcttcttc ttcctcttct tcttcctctt cttcttcctc
     1261 ttcctcttcc ctggcagtat tttcaccaat tctcttgttt aagacggcct gtgcgatttt
     1321 acttgctaat ccattttttt cctccaagat ttcgttttct acctcaaatt cggaggaaga
     1381 agaaggtgca acggatgtcg aagctaagga agactctgac ttacgatgaa aagtagaaga
     1441 gaataatgga atattagaga gggaattgtc actattgacc aatgagacgc catctgaatc
     1501 ttggttatgg tgttcatagg catcgtggtt gtcacgttgg ctgatgatag gtgatgacga
     1561 atctttttct tgtcttttta ttttctgttt tcttagcatt ttcctgcgat tcaattctga
     1621 ggtttttcgg tcttcttcct cttcatcgat gacgtcgaat tcttcacgta gggcggcact
     1681 atcgaaatcg gggaataatt cgcttccaaa gttatttaca acttcgatta ttctttctgc
     1741 cagatttcta cgatcctttt gtagcttttg caagcccttt ggagatgtaa gagataaaac
     1801 cagtggtctc aaagatttga aaccatccat accaatatca cccacgatta aggctgaata
     1861 tgtgactata acacacgaga tgtaagaccc ggaaaaaaca tatattttat tccatggttt
     1921 atgtctgagg taataagtga ttaaaacgga ccaaaagatg taaagcaagg gcgcaaatcc
     1981 catcccaatc aagattttcc acgtggcaat gacatcgtta gcctttattt taactgtaga
     2041 cttggacaaa gcggtacggg ccttttcttg agaaattctc ttggctaata tgaagacagg
     2101 tgagaacata atgatacctg gcatggctaa cgaaaagagg atgcagagcc cgatggatct
     2161 aaaaaaaaca agtccgaggt tttttgcgaa atttaccttt gcctcctcca caaggtgatc
     2221 aggaagatta tagtgtctca aggcggcatt atatgccatt atatctttgg tcaagtccgc
     2281 tattttagga tcgtttctat agaattcgta acctttgacc attcttctgt tcatttccac
     2341 aatcaagggc aacggtaact tggtgctaaa ttgtgtcata tatagtcttc ttatcgtttg
     2401 aaccaccatc aaagtttcat aatcagaaca tgtaacggta acggattgta aacccttcga
     2461 tatggtatct aataactctt tcactgcatc tctgttcgtt tccgagttgt ggtacttggc
     2521 gactagttcc ttcggtattt caatggggtc accaaattca acaaccgctc tcgacctgaa
     2581 cttatgtgga tggaaataat tcataccaca gggaacaatc ttaacattga cgtcaggatg
     2641 cttatccatg caaccaagag ccataatcgc cacacctgct ttcaggggca acaagtttgt
     2701 tctgtcgtgg gacccacctt caggaaagat cccaatgcag ttgttatggg ccaaatgctc
     2761 aaaaactcta tggtaaacgc aagattggtc gactttagcg gcatatttat aagtagtgcc
     2821 ggtgattaaa gcagttttaa tctctggttt ggccattttg aactcttttc ttagggtcaa
     2881 actcgtgtca ctttctatgg actggatttc tccaaatccc attgatttgg gtaacccgat
     2941 gagacccttt ggcatacaat cagtcaagaa atgcgtgtcg tggccgataa ctctcttgta
     3001 gtctgttgga tctacgcgga tagtaccttc tgccggtttc aaattatcct gcggccttac
     3061 cacgcctatg gccatgaaga aactagccaa aaaccctatg gcgggttgct ttaatgagct
     3121 ctccgcaatc aagaaggaca cacgtctgtt gacagatttc ttcacctcgc ccataaggat
     3181 tacagggtcg acgaactggt tagcatgcgg agccgcaaca aagatcaccg gtccctgttg
     3241 cggtaccttg aaactgcccc ggcctctgat ttctctaaag aagcagtgga agatgttgta
     3301 caacagccat gtggcgatgg agtaaagtat cttcttggta gcgctaggct cttggtagat
     3361 gtatgcctga tcagccgacg tcttgacgct tttggcctcg atggaactgt aattcgtagt
     3421 tttctgtgtg ctcttggaag aggcagattt ctccgtgagt tttggtgcag gcattttttt
     3481 ttaccccgat gtaaacgccc ctgtgctctt tttactatat atttaattcc gggcttccta
     3541 cacttaattc actctgttat gaactacaag ccattactaa catcaatctt tccgaatcat
     3601 aaataaactg caactgggaa agaaggcggg cctacttttc tttttcttca tagcgcgtag
     3661 ggcttggcag ccatagttgt tgagagaccg atggatgagg gattgaaaga agggagaaga
     3721 agaatactaa aagaggaatg tagagccggg ttacaggaaa actaactttt cactcagggc
     3781 gccgcatctt ttccgcaggc gcgccatgaa ctggccagca cgttctacgg cgaaaccttc
     3841 gaactggagg cggctcacac atagtagtaa gttgccgcga caggtgacca aaatacagtc
     3901 tttaatatca tcatatttga acatgcccgg tttactgcca ttatctagac gtgcgtctct
     3961 catcacttta cactcatgaa ataatattcg ttttaatact gaattttgag cgtcctttct
     4021 tgccgctgtc gcttctttga acgcgtgcag aggaccgagc gtttcgagtt tgttaagtag
     4081 ttcggcggca gaatcgcgcg cccaatgaat gcgtttatct tctgttgtta tacggtgggc
     4141 gtaactgggt ttgtaagctg ttggcgcttg cacacggttc tgtggtagat ataacctctc
     4201 acgaagcgtt tggcccaaaa gttgggcgcc aagggcgccc aactggtcca tcagtatggc
     4261 agtcctccga ggcaggccct cagaattgaa atctgccgtc gaatcattga ctctcccttt
     4321 tgacagcatt gttgcgatcg ccagtggctc cgtctgcgct acaattgcac catggtcaaa
     4381 ccgatccgga tgcagtgtct gtatagttac accggtgtaa gtgtcaccct ccagcagagc
     4441 tcgctggata ggtgcactgc ctttatgtct gggaagtagc gaaggatgga cgtttagcgc
     4501 ccactagttc tagagcggcc gccaccgcgg tggagctcca attcgcccta tagtgagtcg
     4561 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4621 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4681 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     4741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5161 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     5221 tatttcacac cgcatagatc cgtcgagttc aagagaaaaa aaaagaaaaa gcaaaaagaa
     5281 aaaaggaaag cgcgcctcgt tcagaatgac acgtatagaa tgatgcatta ccttgtcatc
     5341 ttcagtatca tactgttcgt atacatactt actgacattc ataggtatac atatatacac
     5401 atgtatatat atcgtatgct gcagctttaa ataatcggtg tcactacata agaacacctt
     5461 tggtggaggg aacatcgttg gtaccattgg gcgaggtggc ttctcttatg gcaaccgcaa
     5521 gagccttgaa cgcactctca ctacggtgat gatcattctt gcctcgcaga caatcaacgt
     5581 ggagggtaat tctgctagcc tctgcaaagc tttcaagaaa atgcgggatc atctcgcaag
     5641 agagatctcc tactttctcc ctttgcaaac caagttcgac aactgcgtac ggcctgttcg
     5701 aaagatctac caccgctctg gaaagtgcct catccaaagg cgcaaatcct gatccaaacc
     5761 tttttactcc acgcacggcc cctagggcct ctttaaaagc ttgaccgaga gcaatcccgc
     5821 agtcttcagt ggtgtgatgg tcgtctatgt gtaagtcacc aatgcactca acgattagcg
     5881 accagccgga atgcttggcc agagcatgta tcatatggtc cagaaaccct atacctgtgt
     5941 ggacgttaat cacttgcgat tgtgtggcct gttctgctac tgcttctgcc tctttttctg
     6001 ggaagatcga gtgctctatc gctaggggac caccctttaa agagatcgca atctgaatct
     6061 tggtttcatt tgtaatacgc tttactaggg ctttctgctc tgtcatcttt gccttcgttt
     6121 atcttgcctg ctcatttttt agtatattct tcgaagaaat cacattactt tatataatgt
     6181 ataattcatt atgtgataat gccaatcgct aagaaaaaaa aagagtcatc cgctaggtgg
     6241 aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg
     6301 ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa
     6361 tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg
     6421 aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     6481 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6541 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6601 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6661 ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     6721 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     6781 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     6841 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     6901 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     6961 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     7021 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     7081 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     7141 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc
     7201 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7261 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7321 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7381 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7441 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7501 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7561 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7621 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7681 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7741 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7801 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7861 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7921 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7981 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8041 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8101 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8161 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8221 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8341 atcaaaggat cttc
//
