LOCUS       sequence-450706-        8368 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid expressing the SEC18 allele from UWOPS87-2421, under control
            of its native promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8368)
  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 8368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-UWOPS872421
FEATURES             Location/Qualifiers
     source          1..8368
                     /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(4643..4662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4644..4662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4669..4686)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4669..4685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4678..4700)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4826..5281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4913..4932)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5123..5144
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5412..6215)
                     /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(6216..6431)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6455..6474)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6574..6596
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6634..6652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6693..7196
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        7233..7337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7338..7406,7407..8198)
                     /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(7556..7575)
                     /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 cccccccccc ggtgggtgat aattatgaca
     1081 ggcagattcc aatgacagaa taaaaggaaa ggatgccttg gttttgataa ttttataaaa
     1141 tttatcacta aaatatatac catagaggaa agaacttgtt cactaaatcc tttcttattg
     1201 gaagggataa agaataaaaa aacaaggaca tgaaaaattt aaaataatta tgcggattgg
     1261 gtcatcaact caacaagctc gttcacggga tcttcatcgt gccttgcggt ttcaatgttg
     1321 gtcaaggtct ttttaatacc gacattgaag ttaggacagc tcctcgataa ttcattaata
     1381 actttaactc taccagcatc gtcaagaaag tttgattcta tcatgacgtt gttcaattca
     1441 tctaaattgg tcatatttgg aactgctatc tcattgtcga agcaactcaa gatatccatt
     1501 tgttgaagta ccgaataagc tgatgtagta gtcatgatca ataaacgacg gtcttgtggg
     1561 ggtttacgtt tcaatgcaac ctttagcatt tgtaaaatgt tattagagaa tcttggacca
     1621 attggtaccc aatcaactag agtctctaac gaatcaataa caagaatgtt tagtggagat
     1681 ttatacgcat ctctgaaagt gttatcaata taggcaattt ttgcgctttc tgacatgcct
     1741 gacaactcgt tgggagaaat taacctgatg aatgggaatc cagattttaa agcaatttca
     1801 gcggctaaag ctgttttacc gctccctgca gggccgtgga ttaatagaga tactaacctg
     1861 gatttatcac tctcgcgaac ttggcggacg taacgggctc cgttcttcaa tattgagtta
     1921 actcgttcgg aataaagcat cattccacct tccacacatg ttttcaaatc ttcttcacta
     1981 atcccaaaag cgggagtaac atcgttgagt gcatttaaaa agtcttctct tgttacttta
     2041 agttttgcta tatctttagt gttaagtttt gtggcacctt tcccgatgtt gacggttttg
     2101 ttgattgcaa aagaacttgc actcttcact aaaccctcaa tctcagcacc agagaagttt
     2161 tttgttaatg cagctaactc agctaagtta acatcgtcgc tcatcatatt attttccctc
     2221 attttcttcg tctgaatgtc gaaaatttgg agtcttcctt tttcatcggg taaatgaatt
     2281 tcaacttgga cttcaaatct acctggacgc aaaagagcac tgtctattaa atctttacga
     2341 ttggtcatac caataaccaa aatattattc aattgatcaa caccatccat tttagctaac
     2401 aattgattaa ctacattgtc ccctacacca gtaccatcac ctcttgaacc tctctgcttg
     2461 aaaacagaat ccagctcatc gaaaataata atatgtaagg aagattcctc acccttggcc
     2521 ctatattctg cttctgcatc cttaaataaa ttacgaatgt tttcttctga agaaccaacg
     2581 tacttactca aaatttctgg accattgacg attttgggct ctttagcatt cagcattgta
     2641 ccaatctttc ttgcaattaa ggtcttacca gtacctggag gaccgtacaa tagcaaacct
     2701 ttaacatgag aaatacccag tttttctata actgaaggag gaaagattcg acttgcaaac
     2761 gctcttctga aaatcttagt aaactcttta tccaaaccac cgacacccaa atcttcaaac
     2821 ttgaaatccg gtctgatcac agcatttgat cttggtctta atgaatttga tgatttcaaa
     2881 ttaactaaac catctcttcc tttgaaaaaa ttaatttgtg tttgttttgt caaaattccc
     2941 tttgtctcta tcccagttgc aacagcggag gttggttcaa tatcacccaa atcgattgct
     3001 tggacatttc taatttttaa gtcaaagaaa tggccttgga actccatgat aaggtactgg
     3061 gtgggagaaa atatttgaga ttcgtagcaa cgaacaaatt gtttggctaa ctcatcttga
     3121 tcgaataccg tgcttaccgc cttacctcta gctctgaatg agatatctat atctattgaa
     3181 ccaagatacg attgcttacc ggaatacttg aataaatcaa atgcttttgc ttgcacgtct
     3241 tgatttaggg accaaccacc ccaggtacgc tggttaccgt taaatccaat ggttcccggt
     3301 ggaatgtcgt tggagtgtct agttgtgaaa acaaataaat tatcgataat aatataaatg
     3361 ttattaggga aatcacttgg tgagacagca gctacgtttg cgagtgcata ggagttattt
     3421 ggacagtttg acacctttaa atggcgtgta cgggtatcca cgtttgtcat atctggtgga
     3481 gtatgatttg cagcagcttt tccaaaacca ggtatcttga acatactcaa tttataaagg
     3541 ttgaatatac ggtactattt gctgttatct ttgagagaaa caacggtagt acttaaaggc
     3601 tacgaataag ccaatttcat gaatctcgaa atttggtacg cctaacgtat agtgtgatac
     3661 cgaattaacc catgggaaat accttttttt agtgacgcta cgcttgtgta aagagaaaaa
     3721 aatactaggt gacattttag agagggtact gcagaagtat aaagcgtact ctgagtatta
     3781 aagtgcattg aattgaaaat cctacatcaa atacaagtaa atagtacaac ctttataagc
     3841 aaatgtattt aattaaagat aaaatattca actatttagc gcgctcatta ttgattaagg
     3901 cgagggaggc tcaagctaaa tgaactattg ctacccatat 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 tccaccatac cttttgtaat tttcttatac
     4381 acaatgctgt ctgattcttc cggttggatc tttttggcct gcactttggt ttccccgtca
     4441 acagtttgaa actgagtggt cagtaactgt aatggaacag atgaactcaa aactccaaac
     4501 tccttttcta atccaagctc tctaaaacca aagaaatctt caccagtcaa ttcgctggca
     4561 aagtcacctg taaaaaagct ttggctctcg tcttcaagca ctagttctag agcggccgcc
     4621 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     4681 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4741 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4801 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4861 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4921 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4981 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5041 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5101 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5161 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5221 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5281 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     5341 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     5401 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     5461 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     5521 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     5581 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     5641 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     5701 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     5761 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     5821 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     5881 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     5941 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     6001 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     6061 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     6121 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     6181 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     6241 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     6301 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     6361 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     6421 gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6481 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     6541 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     6601 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     6661 ttaatgtcat gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc
     6721 tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt
     6781 tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa
     6841 aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt
     6901 agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat
     6961 cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac
     7021 ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt
     7081 tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt
     7141 aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa
     7201 aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7261 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7321 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7381 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7441 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7501 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7561 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7621 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7681 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7741 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7801 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7861 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7921 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7981 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     8041 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8101 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8161 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8221 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8281 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8341 accccgtaga aaagatcaaa ggatcttc
//
