LOCUS       sequence-358266-        8110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-SWAT compatible vector for C-terminal tagging with SMASh Degron. 
            (Protease Cleavage Site: DEMEECSKHL)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8110)
  AUTHORS   Shively CA, Dong F, Mitra RD
  TITLE     A Suite of New Strain Construction Vectors for Gene Expression 
            Knockdown in Budding Yeast.
  JOURNAL   ACS Synth Biol. 2023 Jan 17. doi: 10.1021/acssynbio.2c00547.
  PUBMED    36650116
REFERENCE   2  (bases 1 to 8110)
  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: "ACS Synth 
            Biol. 2023 Jan 17. doi: 10.1021/acssynbio.2c00547."
COMMENT     Sequence Label: C-SWAT SMASh
FEATURES             Location/Qualifiers
     source          1..8110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             440..1774
                     /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
                     FGGVLSFGVKDLPNADKETDPFELSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     rep_origin      complement(1879..2334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2016..2037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2228..2247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2460..2482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2474..2491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2475..2491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2501..2520
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2501..2519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2544..2560
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2545..2561
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2715..2738
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     gene            3775..5131
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3775..4118
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4119..4928
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4186..4205)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4934..5131
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5337..5357)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5337..5355)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5376..5392)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5376..5392)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5389..5411)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5400..5416
                     /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)."
     promoter        complement(join(5424..5430,5431..5448,5449..5454))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5469..5490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5607..5624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5778..6366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5858..5877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6537..7328,7329..7397))
                     /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     7160..7179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7398..7502)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7539..8042
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8083..8101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcaccat cctcatgaaa actgtgtaac ataataaccg
      241 aagtgtcgaa aaggtggcac cttgtccaat tgaacacgct cgatgaaaaa aataagatat
      301 atataaggtt aagtaaagcg tctgttagaa aggaagtttt tcctttttct tgctctcttg
      361 tcttttcatc tactatttcc ttcgtgtaat acagggtcgt cagatacata gatacaattc
      421 tattaccccc atccatacaa tgccatctca tttcgatact gttcaactac acgccggcca
      481 agagaaccct ggtgacaatg ctcacagatc cagagctgta ccaatttacg ccaccacttc
      541 ttatgttttc gaaaactcta agcatggttc gcaattgttt ggtctagaag ttccaggtta
      601 cgtctattcc cgtttccaaa acccaaccag taatgttttg gaagaaagaa ttgctgcctt
      661 agaaggtggt gctgctgctt tggctgtttc ctccggtcaa gccgctcaaa cccttgccat
      721 ccaaggtttg gcacacactg gtgacaacat cgtttccact tcttacttat acggtggtac
      781 ttataaccag ttcaaaatct cgttcaaaag atttggtatc gaggctagat ttgttgaagg
      841 tgacaatcca gaagaattcg aaaaggtctt tgatgaaaga accaaggctg tttatttgga
      901 aaccattggt aatccaaagt acaatgttcc ggattttgaa aaaattgttg caattgctca
      961 caaacacggt attccagttg tcgttgacaa cacatttggt gccggtggtt acttctgtca
     1021 gccaattaaa tacggtgctg atattgtaac acattctgct accaaatgga ttggtggtca
     1081 tggtactact atcggtggta ttattgttga ctctggtaag ttcccatgga aggactaccc
     1141 agaaaagttc cctcaattct ctcaacctgc cgaaggatat cacggtacta tctacaatga
     1201 agcctacggt aacttggcat acatcgttca tgttagaact gaactattaa gagatttggg
     1261 tccattgatg aacccatttg cctctttctt gctactacaa ggtgttgaaa cattatcttt
     1321 gagagctgaa agacacggtg aaaatgcatt gaagttagcc aaatggttag aacaatcccc
     1381 atacgtatct tgggtttcat accctggttt agcatctcat tctcatcatg aaaatgctaa
     1441 gaagtatcta tctaacggtt tcggtggtgt cttatctttc ggtgtaaaag acttaccaaa
     1501 tgccgacaag gaaactgacc cattcgaact ttctggtgct caagttgttg acaatttaaa
     1561 gcttgcctct aacttggcca atgttggtga tgccaagacc ttagtcattg ctccatactt
     1621 cactacccac aaacaattaa atgacaaaga aaagttggca tctggtgtta ccaaggactt
     1681 aattcgtgtc tctgttggta tcgaatttat tgatgacatt attgcagact tccagcaatc
     1741 ttttgaaact gttttcgctg gccaaaaacc atgagtgtgc gtgatgagtt gtaaaattat
     1801 gtataaacct actttctctc acaagtactg cggtgtgaaa taccgcacag atgcgtaagg
     1861 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1921 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1981 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2041 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2101 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2161 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2221 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2281 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     2341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2461 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     2521 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
     2581 cctgcagcta tattaccctg ttatccctac gtacgctgca ggtcgacggt ggcggttctg
     2641 gcggtggcgg atccgacgaa atggaagaat gtagtaagca cttacctggt gcaggctcat
     2701 ctggcgatat catggactac aaggacgacg atgataaggg ttcttctgga acaggttcag
     2761 gttcaggtac aagtgcaccc attactgctt atgctcagca aacaagaggc ctattgggtt
     2821 gtattattac tagtttgacg ggcagagaca aaaaccaggt agaaggtgaa gtgcagatcg
     2881 tatccacggc tactcagaca ttcttggcta cttgtattaa tggtgtgtgt tgggcagttt
     2941 accacggtgc gggaactaga actattgcct ctcctaaggg tccagtgatc caaatgtata
     3001 ctaacgttga tcaagatttg gtcggctggc cagctccaca agggtcaaga agcctaactc
     3061 cttgcacctg cggttcttca gatctgtact tagtgacacg tcatgcagat gttataccag
     3121 ttagacgtcg tggggacagt agaggttcac tgctgtcacc acgtccaatt agttacctta
     3181 aaggtagttc tggcggtccc ctgttgtgtc ctgcaggtca tgcagtagga ttgtttagag
     3241 ccgccgtctg caccagaggg gttgcaaaag ctgtggattt tattcctgtt gaaaatcttg
     3301 aaacgactat gagatcacct gtctttaccg ataatagttc cccaccagct gtgactctta
     3361 cacatccgat caccaaaatc gatactaaat acattatgac ctgtatgtct gccgatttag
     3421 aagtagtaac atctacatgg gttttagtcg gtggggtttt agctgcccta gccgcatatt
     3481 gcttgtccac aggttgcgtt gttatagttg gcaggatcgt tttgagcgga aaataaagtt
     3541 cttctttgag atatcgattg aacgagctcg aattcatcga tctatattac cctgttatcc
     3601 ctagcgtaac tatgccaaat gacaataaga cacccaatag gtccagcact cccaagttta
     3661 ctaaaaaacc agtaaccccg aatgataaaa ttcctgaaag acggatcccc gggttaatta
     3721 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     3781 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     3841 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     3901 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     3961 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     4021 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     4081 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
     4141 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4201 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     4261 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4321 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4381 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     4441 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4501 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4561 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4621 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     4681 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4741 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4801 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4861 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4921 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     4981 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     5041 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     5101 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     5161 agtttaaacg agctcgaatt ctgatggaaa aagattttca agtgaataag gagataaaac
     5221 cgtattgact ttttatcatt ctacaaaaga ccagtaacga agtgtaccct gctatattac
     5281 cctgttatcc ctagcgtaac tcggccgcca ccgcggtgga gctccagctt ttgttccctt
     5341 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5401 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5461 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5521 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5581 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5641 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5701 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7501 cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt
     7561 ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt
     7621 aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa
     7681 atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt
     7741 gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc
     7801 gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt
     7861 tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac
     7921 cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa acacagagta
     7981 aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga
     8041 tccgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8101 ccctttcgtc
//
