LOCUS       sequence-450713-        7511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the SEC22 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 7511)
  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 7511)
  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: pRS315-SEC22-NCYC110
FEATURES             Location/Qualifiers
     source          1..7511
                     /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(2662..2681)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2663..2681)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2688..2705)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2688..2704)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2697..2719)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2845..3300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2932..2951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3142..3163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3601..4008
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4009..5103
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(5597..5616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5716..5738
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5776..5794)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5836..6339
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6376..6480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6481..6549,6550..7341)
                     /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(6699..6718)
                     /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 gcccccccaa agactcccta aaaagtgcag
     1081 gtgcggcggg tcacccgcca gcttgcctga aaaatacgta attatgtcaa ataaattgcc
     1141 atactatact aatactgaac tagactatgt tatacacttg gaccaaattg atcgggattt
     1201 gtgatgtggg atgatggggt gacgtctatt tgaggaagat ccaccagaag agaaagacga
     1261 aaaagaaagc gacaatgaca ataggagcat attggctgat caagagatcg aagttgatct
     1321 tttgcgcgga ctttctgtac cttttggatg tttctttcaa agaagaactc atgtcactca
     1381 ttttatcgag agaatctccc ctgtaaagta ggtcttcgat gttcttggac atgatttgct
     1441 tgacacctac aagctcttgg tttagttggt ccaaattgtc ctgaactttc ttatcactgt
     1501 atgacttttt tgtcatctgt agaaagttat caaagtttac aaactggtat ggtcttacgc
     1561 taggcttggg gtactcatta gcaaatgagt gttcaaattc ttgcgctata tcgttaaggt
     1621 atgagaatgc taagtttctt gggtatccag attcgcaaat gacgaagtag tacaccatgg
     1681 atttcttcaa ataatggatc tcaaaggagc cactttccaa agtagcctcc gtggcagact
     1741 gtggtgtcaa tctggaaacg acgattttca ccttttgctt ttgttcaaat aatgagggat
     1801 cattttcgtt gtccacagac gtacaaagag gcagcccatc ttctctgtag attagtgttg
     1861 actttatcat ccctacttaa ttgtgagtgt gtaacggggt gtcactgtca gggtttttgt
     1921 tggttttggg taataatagg ttgtagatat attatattac atgggctcta cttgtttatc
     1981 ttctatggta gaaaaagacc gaaaaatgaa aacaaaaagc gttgaggatt tattgctaaa
     2041 aaaaaattcg tttatgacgt ctcttattag ataaatcaag tgataactag agaaatttat
     2101 ttaattatat atgctagtac aacacgtttg gttgaaaatt agacctcaaa ccacgacaaa
     2161 cttctgtctt atatcgaaga aagttgggaa actctcccgc aggtccagat ttggtaaatg
     2221 gtcttgtagg ttactgatat taagaacgta gttagagtcg tcctctaatt gagcaaatcc
     2281 caatttcttg cattgtggta gttcattcca tgaagtgaac tctatacaac tcatatcatt
     2341 tgacgcttgt ttttggtgtt gggtggtgtc gcaaagggta ttgttgatga acagttcctg
     2401 cagatttggt aggtaaaatc cctggattcg gatggacgaa catttttcaa tacgtgtgat
     2461 tttgacagtg agcgatctta aagagttgtt cttcaagcaa atcattggcc tctttaaatg
     2521 gtgtctttgt ttgaaggtat catcgctaaa tgaagaagta aaggggatcc agttgattat
     2581 ttgagttttt tctaccccct caggtatcaa cttgactcac tagttctaga gcggccgcca
     2641 ccgcggtgga gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     2701 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2761 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2821 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2881 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2941 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3001 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3061 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3121 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3181 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3241 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3301 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatcgaccgg
     3361 tcgaggagaa cttctagtat atctacatac ctaatattat tgccttatta aaaatggaat
     3421 cccaacaatt acatcaaaat ccacattctc ttcaaaatca attgtcctgt acttccttgt
     3481 tcatgtgtgt tcaaaaacgt tatatttata ggataattat actctatttc tcaacaagta
     3541 attggttgtt tggccgagcg gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt
     3601 aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta
     3661 tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat caaattcgat
     3721 gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt ttcaactgaa
     3781 aaattgggag aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat agaatagaga
     3841 agcgttcatg actaaatgct tgcatcacaa tacttgaagt tgacaatatt atttaaggac
     3901 ctattgtttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt tcttaccttt
     3961 tacatttcag caatatatat atatatattt caaggatata ccattctaat gtctgcccct
     4021 aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc cgaagccatt
     4081 aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt cgaaaatcat
     4141 ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga ggcgctggaa
     4201 gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc
     4261 ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca attgtacgcc
     4321 aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc aatcaagcca
     4381 caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg tatttacttt
     4441 ggtaagagaa aggaagacga tggtgatggt gtcgcttggg atagtgaaca atacaccgtt
     4501 ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca tgagccacca
     4561 ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt atggagaaaa
     4621 actgtggagg aaaccatcaa gaacgaattc cctacattga aggttcaaca tcaattgatt
     4681 gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat tataatcacc
     4741 agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg ttccttgggt
     4801 ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt tggtttgtac
     4861 gaaccatgcc acggttctgc tccagatttg ccaaagaata aggtcaaccc tatcgccact
     4921 atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga aggtaaggcc
     4981 attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga tttaggtggt
     5041 tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa aatccttgct
     5101 taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga aattcataat
     5161 agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa ctatatacgc
     5221 aatctacata catttatcaa gaaggagaaa aaggaggatg taaaggaata caggtaagca
     5281 aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac attaatattg
     5341 acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactatgatt
     5401 cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca
     5461 ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc atttcccata
     5521 atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg acgtagtcga
     5581 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5641 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5701 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5761 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5821 tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
     5881 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
     5941 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
     6001 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
     6061 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
     6121 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
     6181 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
     6241 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
     6301 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
     6361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7501 aaaggatctt c
//
