LOCUS       sequence-204894-        7783 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 7783)
  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 7783)
  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..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     216..236
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        218..236
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2627..2800
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2801..2974
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      3010..3197
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3065..3089)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3220..3236)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3221..3237)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3261..3280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3262..3280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3290..3307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3290..3306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3299..3321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3447..3902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3534..3553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3744..3765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4033..4836)
                     /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(4837..5052)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5076..5095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5195..5217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5255..5273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5314..5817
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5854..5958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5959..6027,6028..6819)
                     /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(6177..6196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6990..7578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7479..7498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7732..7749
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      241 aaaagctgga gctcttgcat ggaggcgcaa ttggcaaaga cgtaaagctc ttgtcatcaa
      301 ttagatctaa tttggaaaca gattggtaca atttggatgg aattttaggc ttggcaccta
      361 tcaattgtga gcgacgcttt cttacattca ataactgttg gttctgattt tcactttgac
      421 tattgccttg gtatggagaa tttctctgca atggttttgc ctgcttcgaa ggagtgctgt
      481 tactatcgaa gataaattcc ttcttttcaa attgccattg taacatggag ttattctcta
      541 tatcctggac gactggagga gtcaagggcc tggaaagtac ggatcctgta gaaatatcac
      601 tggcaattat actgagttta ttgttggtgt tgctattcat atttgagctg gtattggacg
      661 aaagcaagga taatgagttt gtgtctactg actccaacac cctaggctgg atttgatctt
      721 tctccatcct ttaattttaa ccttttaatt agtggttgga tcaagttttc gagactattc
      781 caatctgtga cttgttggat aaatagtttt tgactcgttt agtataatcc ttttttctaa
      841 aagtgcttag agttctctaa gatgttcttg tttacaatgt gagcgattta ggaaatttcc
      901 taaaaatggc cgaggcggcg ctagcattct acgaaaggtg agataacgct tcgttatcga
      961 aaaatgtcag gggacagggg ttatataaga acgaaaattg tcatcctgca ttttttcttt
     1021 aaaacagcta tacaaaaagt gataccgaca tacaatgaag tatctaaact tagttttcgt
     1081 gcttcagctt cttattagca tcaaatacgc ctcattcggc cgagcctttt ctctttttga
     1141 agatgatacc acctttgcca atttggataa acagctaaag cttccacaga atacacagca
     1201 aacccttaaa ttggaccgtt tgaatcacga tgatccgctg tttacaactt ttatttcttc
     1261 tgtggacaca gattacagtt tgagacttag aacagtagat ccttctaaac taggaattga
     1321 caccgtaaaa caatggtcgg gttacatgga ctataaggat tccaaacact ttttttactg
     1381 gttttttgaa agtaggaacg atcctgctaa cgacccaatt attctttggt taaatggtgg
     1441 acctggttgt tcctcgttta ctgggttgct atttgaacta ggcccctcat caattggcgc
     1501 cgatatgaaa ccaatccaca atccctattc ttggaataat aacgcttcaa tgatcttctt
     1561 agaacagcca ctcggagtcg gcttttccta tggtgatgaa aaagtctcct ctacaaaatt
     1621 agcaggcaaa gatgcgtaca ttttcctgga attgtttttt gaagcttttc ctcatttacg
     1681 ctccaacgat ttccacattg caggcgaatc ctatgcagga cattatatcc ctcaaattgc
     1741 acatgagatc gttgtcaaga accctgaaag aacgttcaat ttaacttcag ttatgattgg
     1801 taatggtatc acagaccctt tgattcaagc agattattat gaaccaatgg catgcgggaa
     1861 agggggctat caccctgttc tctcatcaga agaatgtgag aaaatgagta aagctgcagg
     1921 tcgttgtcgt aggttgaaca agttatgtta tgcttctaaa tcaagtttac catgcatagt
     1981 cgccactgct tactgtgact ctgcactttt ggaaccgtac attaacacag gactcaacgt
     2041 ctatgacatt agagggccct gtgaagataa tagtactgat ggtatgtgtt atacaggtct
     2101 ccgctatgtc gaccagtata tgaattttcc tgaagttcaa gaaacgctag ggtccgacgt
     2161 gcataattat tctggctgtg ataatgacgt gttcaccgga tttttgttta cgggcgatgg
     2221 aagtaaacca tttcaacaat atattgctga attattaaat cacaacattc cggtattaat
     2281 atatgcgggt gataaggatt atatttgtaa ttggctggga aaccatgctt ggtccaatga
     2341 gttggaatgg atcaataaac gtaggtatca gagaaggatg ttaagaccat gggtcagtaa
     2401 agaaacaggt gaagagttgg gacaagtcaa gaactatggc cctttcacct ttttgagaat
     2461 atacgatgcc ggtcatatgg tgccctatga tcaaccggag gcaagtttgg aaatggtcaa
     2521 cagttggatt tccggtaatc gtgctttttc ggatctttcc accttggaaa atgctagttc
     2581 tagacggatc cccgggttaa ttaagcttgc gcaacacgat gaagccgtgg acaacaaatt
     2641 caacaaagaa caacaaaacg cgttctatga gatcttacat ttacctaact taaacgaaga
     2701 acaacgaaac gccttcatcc aaagcttaaa agatgaccca agccaaagcg ctaacctttt
     2761 agcagaagct aaaaagctaa atgatgctca ggcgccgaaa gtagacaaca aattcaacaa
     2821 agaacaacaa aacgcgttct atgagatctt acatttacct aacttaaacg aagaacaacg
     2881 aaacgccgtc atccaaagtt taaaagatga cccaagccaa agcgctaacc ttttagcaga
     2941 agctaaaaag ctaaatgatg ctcaggcgcc gaaagtagac gcgaattaag gcgcgccact
     3001 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     3061 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     3121 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
     3181 gaccacacct ctaccggcag aattcgatat caagcttatc gataccgtcg acctcgaggg
     3241 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     3301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3421 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     3481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3601 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3661 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3721 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3781 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3841 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     3901 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa
     3961 taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta
     4021 taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt
     4081 tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac
     4141 aataataatg tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa
     4201 tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc
     4261 atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt
     4321 cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa
     4381 ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa
     4441 accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc
     4501 tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt
     4561 tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt
     4621 gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg
     4681 acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca
     4741 caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg
     4801 agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt
     4861 ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca
     4921 tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga
     4981 gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa
     5041 tgatgaattg aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5101 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5161 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5221 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5281 gttaatgtca tgataataat ggtttcttag acggatcgct tgcctgtaac ttacacgcgc
     5341 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
     5401 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
     5461 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
     5521 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
     5581 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
     5641 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
     5701 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
     5761 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
     5821 aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5881 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5941 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6001 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6061 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6121 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6181 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6241 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6301 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6361 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6421 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6481 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6541 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6601 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6661 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6721 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6781 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6841 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6901 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6961 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7021 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7081 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7141 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7201 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7261 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7321 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7381 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7441 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7501 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7561 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7621 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7681 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7741 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctc
//
