LOCUS       sequence-201579-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Atg42/Ybr139w with a C-terminal PA tag in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  AUTHORS   Parzych KR, Ariosa A, Mari M, Klionsky DJ
  TITLE     A newly characterized vacuolar serine carboxypeptidase, 
            Atg42/Ybr139w, is required for normal vacuole function and the 
            terminal steps of autophagy in the yeast Saccharomyces cerevisiae.
  JOURNAL   Mol Biol Cell. 2018 May 1;29(9):1089-1099. doi: 
            10.1091/mbc.E17-08-0516. Epub 2018 Mar 22.
  PUBMED    29514932
REFERENCE   2  (bases 1 to 7795)
  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"; doi: 
            "10.1091/mbc.E17-08-0516"; journalName: "Mol Biol Cell"; date: 
            "2018-05-1- 1"; volume: "29"; issue: "9"; pages: "1089-1099"
COMMENT     Sequence Label: pKP105 (pRS416-YBR139Wp-YBR139W-PA-ADH1t)
FEATURES             Location/Qualifiers
     source          1..7795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             5166..5339
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             5340..5513
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      5549..5736
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5604..5628)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5759..5775)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5760..5776)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5800..5819)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5801..5819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5829..5846)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5829..5845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5838..5860)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5990..6445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6077..6096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6287..6308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6576..7379)
                     /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(7380..7600)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7626..7645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7745..7767
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcttgcatg gaggcgcaat tggcaaagac
     2821 gtaaagctct tgtcatcaat tagatctaat ttggaaacag attggtacaa tttggatgga
     2881 attttaggct tggcacctat caattgtgag cgacgctttc ttacattcaa taactgttgg
     2941 ttctgatttt cactttgact attgccttgg tatggagaat ttctctgcaa tggttttgcc
     3001 tgcttcgaag gagtgctgtt actatcgaag ataaattcct tcttttcaaa ttgccattgt
     3061 aacatggagt tattctctat atcctggacg actggaggag tcaagggcct ggaaagtacg
     3121 gatcctgtag aaatatcact ggcaattata ctgagtttat tgttggtgtt gctattcata
     3181 tttgagctgg tattggacga aagcaaggat aatgagtttg tgtctactga ctccaacacc
     3241 ctaggctgga tttgatcttt ctccatcctt taattttaac cttttaatta gtggttggat
     3301 caagttttcg agactattcc aatctgtgac ttgttggata aatagttttt gactcgttta
     3361 gtataatcct tttttctaaa agtgcttaga gttctctaag atgttcttgt ttacaatgtg
     3421 agcgatttag gaaatttcct aaaaatggcc gaggcggcgc tagcattcta cgaaaggtga
     3481 gataacgctt cgttatcgaa aaatgtcagg ggacaggggt tatataagaa cgaaaattgt
     3541 catcctgcat tttttcttta aaacagctat acaaaaagtg ataccgacat acaatgaagt
     3601 atctaaactt agttttcgtg cttcagcttc ttattagcat caaatacgcc tcattcggcc
     3661 gagccttttc tctttttgaa gatgatacca cctttgccaa tttggataaa cagctaaagc
     3721 ttccacagaa tacacagcaa acccttaaat tggaccgttt gaatcacgat gatccgctgt
     3781 ttacaacttt tatttcttct gtggacacag attacagttt gagacttaga acagtagatc
     3841 cttctaaact aggaattgac accgtaaaac aatggtcggg ttacatggac tataaggatt
     3901 ccaaacactt tttttactgg ttttttgaaa gtaggaacga tcctgctaac gacccaatta
     3961 ttctttggtt aaatggtgga cctggttgtt cctcgtttac tgggttgcta tttgaactag
     4021 gcccctcatc aattggcgcc gatatgaaac caatccacaa tccctattct tggaataata
     4081 acgcttcaat gatcttctta gaacagccac tcggagtcgg cttttcctat ggtgatgaaa
     4141 aagtctcctc tacaaaatta gcaggcaaag atgcgtacat tttcctggaa ttgttttttg
     4201 aagcttttcc tcatttacgc tccaacgatt tccacattgc aggcgaatcc tatgcaggac
     4261 attatatccc tcaaattgca catgagatcg ttgtcaagaa ccctgaaaga acgttcaatt
     4321 taacttcagt tatgattggt aatggtatca cagacccttt gattcaagca gattattatg
     4381 aaccaatggc atgcgggaaa gggggctatc accctgttct ctcatcagaa gaatgtgaga
     4441 aaatgagtaa agctgcaggt cgttgtcgta ggttgaacaa gttatgttat gcttctaaat
     4501 caagtttacc atgcatagtc gccactgctt actgtgactc tgcacttttg gaaccgtaca
     4561 ttaacacagg actcaacgtc tatgacatta gagggccctg tgaagataat agtactgatg
     4621 gtatgtgtta tacaggtctc cgctatgtcg accagtatat gaattttcct gaagttcaag
     4681 aaacgctagg gtccgacgtg cataattatt ctggctgtga taatgacgtg ttcaccggat
     4741 ttttgtttac gggcgatgga agtaaaccat ttcaacaata tattgctgaa ttattaaatc
     4801 acaacattcc ggtattaata tatgcgggtg ataaggatta tatttgtaat tggctgggaa
     4861 accatgcttg gtccaatgag ttggaatgga tcaataaacg taggtatcag agaaggatgt
     4921 taagaccatg ggtcagtaaa gaaacaggtg aagagttggg acaagtcaag aactatggcc
     4981 ctttcacctt tttgagaata tacgatgccg gtcatatggt gccctatgat caaccggagg
     5041 caagtttgga aatggtcaac agttggattt ccggtaatcg tgctttttcg gatctttcca
     5101 ccttggaaaa tgctagttct agacggatcc ccgggttaat taagcttgcg caacacgatg
     5161 aagccgtgga caacaaattc aacaaagaac aacaaaacgc gttctatgag atcttacatt
     5221 tacctaactt aaacgaagaa caacgaaacg ccttcatcca aagcttaaaa gatgacccaa
     5281 gccaaagcgc taacctttta gcagaagcta aaaagctaaa tgatgctcag gcgccgaaag
     5341 tagacaacaa attcaacaaa gaacaacaaa acgcgttcta tgagatctta catttaccta
     5401 acttaaacga agaacaacga aacgccgtca tccaaagttt aaaagatgac ccaagccaaa
     5461 gcgctaacct tttagcagaa gctaaaaagc taaatgatgc tcaggcgccg aaagtagacg
     5521 cgaattaagg cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat
     5581 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     5641 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     5701 gtatgaggtc gctcttattg accacacctc taccggcaga attcgatatc aagcttatcg
     5761 ataccgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
     5821 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5881 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5941 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt
     6001 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6061 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6121 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6181 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6241 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6301 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6361 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6421 aacaaaatat taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     6481 gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag
     6541 tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc
     6601 aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc
     6661 tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc
     6721 acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc
     6781 ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg
     6841 ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta
     6901 gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt
     6961 acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca
     7021 ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact
     7081 gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat
     7141 aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt
     7201 gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg
     7261 gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc
     7321 ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg
     7381 atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa
     7441 tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc
     7501 cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat
     7561 caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact
     7621 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7681 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7741 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
