LOCUS       sequence-450712-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the SEC18 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 8370)
  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 8370)
  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-L1374
FEATURES             Location/Qualifiers
     source          1..8370
                     /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(4645..4664)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4646..4664)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4671..4688)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4671..4687)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4680..4702)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4828..5283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4915..4934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5125..5146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5414..6217)
                     /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(6218..6433)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6457..6476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6576..6598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6636..6654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6695..7198
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7235..7339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7340..7408,7409..8200)
                     /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(7558..7577)
                     /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 ttttgttaat 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 agtaccatca cctcttgaac ctctctgctt
     2461 gaaaacagaa tccagctcat cgaaaataat aatatgtaag gaagattcct cacccttggc
     2521 cctatattct gcttctgcat ccttaaataa attacgaatg ttttcttctg aagaaccaac
     2581 gtacttactc aaaatttctg gaccattgac gattttgggc tctttagcat 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 ggtcaaagaa atggccttgg aactccatga taaggtactg
     3061 ggtgggagaa aatatttgag attcgtagca acgaacaaat tgtttggcta actcatcttg
     3121 gtcgaatacc 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 acgtttgtca tatctggtgg
     3481 agtatgattt gcagcagctt ttccaaaacc aggtatcttg aacatactca atttataaag
     3541 gttgaatata 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 tctgagtgtt
     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 gtggaagtga ttgaggcttt 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 caccagtcag ttcgctggca
     4561 aagtcacctg taaaaaagct ttggctctcg tcttcaaagc actagttcta gagcggccgc
     4621 ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     4681 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4741 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4801 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     4861 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4921 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4981 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5041 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5101 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5161 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5221 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5281 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta
     5341 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     5401 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     5461 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     5521 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     5581 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     5641 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     5701 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     5761 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     5821 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     5881 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     5941 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     6001 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     6061 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     6121 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     6181 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     6241 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     6301 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     6361 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
     6421 atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata
     6481 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6541 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6601 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6661 ggttaatgtc atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg
     6721 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
     6781 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
     6841 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
     6901 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
     6961 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
     7021 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
     7081 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt
     7141 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga
     7201 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7561 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7861 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8341 agaccccgta gaaaagatca aaggatcttc
//
