LOCUS       sequence-450711-        8369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the SEC18 allele from S288C, under control of its
            native promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8369)
  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 8369)
  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: pRS316-SEC18-S288C
FEATURES             Location/Qualifiers
     source          1..8369
                     /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(4644..4663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4645..4663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4670..4687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4670..4686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4679..4701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4827..5282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4914..4933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5124..5145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5413..6216)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6217..6432)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6456..6475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6575..6597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6635..6653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6694..7197
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7234..7338
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7339..7407,7408..8199)
                     /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(7557..7576)
                     /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 gccccccccc cggtgggtga taattatgac
     1081 aggcagattc caatgacaga ataaaaggaa aggatgcctt ggttttgata attttataaa
     1141 atttatcact aaaatatata ccatagagga aagaacttgt tcactagatc ctttctgatt
     1201 ggaagggata aagaataaaa atacaaggac atgaaaaatt tgaaataatt atgcggattg
     1261 ggtcatcaac tcaacaagct cgttcacggg atcttcatcg tgccttgcgg tttcaatgtt
     1321 ggtcaaggtc tttttaatac cgacattgaa gttaggacag ctccttgata attcattaat
     1381 aactttaact ctaccagcat cgtcaagaaa gtttgattct atcatgacgt tgttcaattc
     1441 atctaaattg gtcatatttg gaactgctat ctcattgtcg aagcaactca agatatccat
     1501 ttgttgaagt accgaataag ctgatgtagt agtcatgatc aataaacgac ggtcttgtgg
     1561 gggtttacgc ttcaatgcaa cctttagcat ttgtaaaatg ttattagaga atcttggacc
     1621 aattggtacc caatcaacta gagtctctaa cgaatcaata acaagaatgt ttagtggaga
     1681 tttatacgca tctctgaaag tgttatcaat ataggcaatt tttgcgcttt ctgacatgcc
     1741 tgacaactcg ttgggagaaa ttaacctgat gaatgggaat ccagatttta aagcaatttc
     1801 agcggctaaa gctgttttac cggaccctgc agggccgtgg attaatagag atactaacct
     1861 ggatttatca ctctcgcgaa cttggcggac gtaacgggct ccgttcttca atattgagtt
     1921 aactcgttcg gaataaagca tcattccacc ttccacacat gttttcaaat cttcttcact
     1981 aatcccaaaa gcgggagtaa catcgttgag tgcatttaaa aagtcttctc ttgttacttt
     2041 aagttttgct atatctttag tgttaagttt tgtggcacct ttcccgatgt tgacggtttt
     2101 gttgattgca aaagaacttg cactcttcac taaaccctca atctcagcac cagagaagtt
     2161 ttttgttaac gcagctaact cagctaagtt aacatcgtcg ctcatcatat tattttccct
     2221 cattttcttc gtctgaatgt cgaaaatttg gagtcttcct ttttcatcgg gtaaatgaat
     2281 ttcaacttgg acttcaaatc tacctggacg caaaagagca ctgtctatta aatctttacg
     2341 attggtcata ccaataacca aaatattatt caattgatca acaccatcca ttttagctaa
     2401 caattgatta actacattgt cccctacacc ggtaccatca cctcttgaac ctctctgctt
     2461 gaaaacagaa tccagctcat cgaaaataat aatatgtaag gaagattcct cacccttggc
     2521 cctatattct gcttctgcat ccttaaataa attacgaatg ttttcttctg aagaaccaac
     2581 gtacttactc aaaatttctg gaccattgac gattttgggc tctttggcat tcagcattgt
     2641 accaatcttt cttgcaatta aggtcttacc agtacctgga ggaccgtaca atagcaaacc
     2701 tttaacatga gaaataccca gtttttctat aactgaagga ggaaagattc gacttgcaaa
     2761 cgctcttctg aaaattttag taaactcttt atccaaacca ccgacaccca aatcttcaaa
     2821 cttgaaatcc ggtctgatca cagcatttga tcttggtctt aatgaatttg atgatttcaa
     2881 attaactaaa ccatctcttc ctttgaaaaa attaatttgt gtttgttttg tcaaaattcc
     2941 ctttgtctct atcccagttg caacagcgga ggttggttca atatcaccca aatcgattgc
     3001 ttggacattt ctaattttta agtcaaagaa atggccttgg aactccatga taaggtactg
     3061 ggtgggagaa aatatttgag attcgtagca acgaacaaat tgtttggcta actcatcttg
     3121 atcgaatacc gtgcttaccg ccttacctct agctctgaat gagatatcta tatctattga
     3181 accaagatac gattgcttac cggaatactt gaataaatca aatgcttttg cttgcacgtc
     3241 ttgatttagg gaccaaccac cccaggtacg ctggttaccg ttaaatccaa tggttcccgg
     3301 tggaatgtcg ttggagtgtc tagttgtgaa aacaaataaa ttatcgataa taatataaat
     3361 gttattaggg aaatcatttg gtgagacagc agctacgttt gcgagtgcat aggagttatt
     3421 tggacagttt gacaccttta aatggcgtgt acgggtatcc atgtttgtca tatctggtgg
     3481 agtatgattt gcagcagctt ttccaaaacc aggtatcttg aacatactca atttataaag
     3541 gctgaatata cggtactatt tgctcttatc tttgagagaa acaacggtag tacttaaagg
     3601 ctacgaataa gccaatttca tgaatctcga aatttggtac gcctaacgta tagtgtgata
     3661 ccgaattaac ccatgggaaa tacctttttt tagtgacgct acgcttgtgt aaagagaaaa
     3721 aaatactagg tgacatttta gagagggtac tgcagaagta taaagcgtac tctgagtatt
     3781 aaagtgcatt gaattggaat cctacatcaa ctacaagtaa atagtacaac ctttataagc
     3841 aaatgtattt aattaaagat aaaatattca actatttagc gcgctcatta ttgattaagg
     3901 cgaggaaggc tcaagctaaa tgaactattg ctgcccatat tggtatcatc aaaagcatca
     3961 tccatatcca aaggatctaa aggttgaggc gttcgcaaaa ataaggaaga atctccatca
     4021 ttttctgcac ccactttggc gtttacagtt gttgttggat ccgcttttac gtcgttttcc
     4081 aagtcttctt ctggaagtat aaacgcgctt gcgatcggtt tctttttgta tgtggtactt
     4141 atcttaccag taggaggtaa tcttgccttc gttgcggttt tctttgagac aaactgatcc
     4201 tcttctagta aagcgaaaga attgtcttct gtggaagtca ttgaggtttt atcatttgca
     4261 gatgacttgc tttgctttgc tatataggcc ttggaacgtt cataatcttt ttgtaatagg
     4321 ggaagtagag tattccagaa tgaaccagca tccagcatac cttttgtaat tttcttatac
     4381 acaatgctgt ctgattcttc cggttggatc tttttggcct gcactttggt ttccccgtca
     4441 acagtttgaa actgagtagt cagtaactgt aatggaacag atgaactcaa aactccaaac
     4501 tccttttcta atccaagctc tctaaaacca aagaagtctt caccagtcaa ttcgctggca
     4561 aagtcacctg taaaaaagct ttggctctcg tcttcaaagc actagttcta gagcggccgc
     4621 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     4681 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4741 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4801 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4981 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5041 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5101 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5161 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5221 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5281 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa
     5341 taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta
     5401 taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt
     5461 tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac
     5521 aataataatg tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa
     5581 tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc
     5641 atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt
     5701 cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa
     5761 ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa
     5821 accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc
     5881 tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt
     5941 tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt
     6001 gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg
     6061 acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca
     6121 caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg
     6181 agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt
     6241 ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca
     6301 tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga
     6361 gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa
     6421 tgatgaattg aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6481 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6541 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6601 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     6661 gttaatgtca tgataataat ggtttcttag acggatcgct tgcctgtaac ttacacgcgc
     6721 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
     6781 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
     6841 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
     6901 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
     6961 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
     7021 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
     7081 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
     7141 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
     7201 aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7321 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7381 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7441 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7501 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7561 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7621 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7681 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7741 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7801 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7861 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7921 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7981 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8041 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8101 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8161 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8341 gaccccgtag aaaagatcaa aggatcttc
//
