LOCUS       sequence-109819-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of S. cerevisiae MET25 cloned into pFA6a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  TITLE     Houseley Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4937)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFa6a MET25
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             279..1613
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     complement(2512..2531)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2513..2531)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2618..2635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2789..3377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2869..2888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3548..4339,4340..4408))
                     /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     4171..4190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4409..4513)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4581..4599
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4637..4659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4759..4778
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4858..4876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4858..4875
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cgcgcctggt ggttggcaaa tgactaatta agttagtcaa ggcgccatcc tcatgaaaac
       61 tgtgtaacat aataaccgaa gtgtcgaaaa ggtggcacct tgtccaattg aacacgctcg
      121 atgaaaaaaa taagatatat ataaggttaa gtaaagcgtc tgttagaaag gaagtttttc
      181 ctttttcttg ctctcttgtc ttttcatcta ctatttcctt cgtgtaatac agggtcgtca
      241 gatacataga tacaattcta ttacccccca tccatacaat gccatctcat ttcgatactg
      301 ttcaactaca cgccggccaa gagaaccctg gtgacaatgc tcacagatcc agagctgtac
      361 caatttacgc caccacttct tatgttttcg aaaactctaa gcatggttcg caattgtttg
      421 gtctagaagt tccaggttac gtctattccc gtttccaaaa cccaaccagt aatgttttgg
      481 aagaaagaat tgctgcttta gaaggtggtg ctgctgcttt ggctgtttcc tccggtcaag
      541 ccgctcaaac ccttgccatc caaggtttgg cacacactgg tgacaacatc gtttccactt
      601 cttacttata cggtggtact tataaccagt tcaaaatctc gttcaaaaga tttggtatcg
      661 aggctagatt tgttgaaggt gacaatccag aagaattcga aaaggtcttt gatgaaagaa
      721 ccaaggctgt ttatttggaa accattggta atccaaagta caatgttccg gattttgaaa
      781 aaattgttgc aattgctcac aaacacggta ttccagttgt cgttgacaac acatttggtg
      841 ccggtggtta cttctgtcag ccaattaaat acggtgctga tattgtaaca cattctgcta
      901 ccaaatggat tggtggtcat ggtactacta tcggtggtat tattgttgac tctggtaagt
      961 tcccatggaa ggactaccca gaaaagttcc ctcaattctc tcaacctgcc gaaggatatc
     1021 acggtactat ctacaatgaa gcctacggta acttggcata catcgttcat gttagaactg
     1081 aactattaag agatttgggt ccattgatga acccatttgc ctctttcttg ctactacaag
     1141 gtgttgaaac attatctttg agagctgaaa gacacggtga aaatgcattg aagttagcca
     1201 aatggttaga acaatcccca tacgtatctt gggtttcata ccctggttta gcatctcatt
     1261 ctcatcatga aaatgctaag aagtatctat ctaacggttt cggtggtgtc ttatctttcg
     1321 gtgtaaaaga cttaccaaat gccgacaagg aaactgaccc attcaaactt tctggtgctc
     1381 aagttgttga caatttaaag cttgcctcta acttggccaa tgttggtgat gccaagacct
     1441 tagtcattgc tccatacttc actacccaca aacaattaaa tgacaaagaa aagttggcat
     1501 ctggtgttac caaggactta attcgtgtct ctgttggtat cgaatttatt gatgacatta
     1561 ttgcagactt ccagcaatct tttgaaactg ttttcgctgg ccaaaaacca tgagtgtgcg
     1621 taatgagttg taaaattatg tataaaccta ctttctctca caagtactat acttttataa
     1681 aacgaacttt attgaaatga atatcctttt tttcccttgt tacatgtcgt gactcgtact
     1741 ttgaacctaa attgttctaa catcaaagaa cagtgttaat tcgcagtcga gaagaaaaat
     1801 atggtgaaca agactcatct acttcatgag actactttac gcctcctata aagctgtcac
     1861 actggataaa tttattgtag gaccaagtta caaaagagga tgatggaggt ttctttacaa
     1921 taaagaagca catgtgtgtt aacgttttta gtatttgctt gttatgtaaa tcaggaaaac
     1981 ttcgcgggat ttggttggat gctactttcc atacaataaa tattatagat ctaaaaagcc
     2041 aaattacaag taaagattag taaagctgtt ggaattccat cgttgataaa aatgttagtt
     2101 attaaatata aaagtcagaa taggtgaact tggatttaat tgttggcatt tcgttgctgc
     2161 tagaggccat aatattagat agccaggaca tactagttct cctcgtggta taggaatcca
     2221 taaaatggaa ttggtgattc tatgtgatat attcacattc ttactacatt atcaatcctt
     2281 gcacttcagc ttcctctaac ctcgatgaca tcttctcata acttatgtca tcatctaacg
     2341 ccgtctatta taatatattg atagtataag tattagttga tagacaatag tggattttta
     2401 ttccaacagt gtctttgttc gtctcagata tagtcggagt ttaaacgagc tcgaattcat
     2461 cgatgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     2521 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     2581 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     3001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3841 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4141 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4501 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4561 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4621 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4681 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4741 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca
     4801 tattgtcgtt agaacgcggc tcaattaata cataacctta tgtatcatac acatacgatt
     4861 taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag gtcgacggat
     4921 ccccgggtta attaagg
//
