LOCUS       sequence-204895-        7783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Atg42/Ybr139w(N163,242Q) 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: pKP110 [pRS416-YBR139Wp-YBR139W(N163,242Q)-PA-...
FEATURES             Location/Qualifiers
     source          1..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..30
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    147..168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(183..188,189..206,207..213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..237
                     /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     226..248
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     245..261
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     245..261
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     280..300
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        282..300
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2691..2864
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2865..3038
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      3074..3261
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3129..3153)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3284..3300)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3285..3301)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3325..3344)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3326..3344)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3354..3371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3354..3370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3363..3385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3511..3966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3598..3617)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3808..3829
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4097..4900)
                     /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(4901..5116)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5140..5159)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5259..5281
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5319..5337)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5378..5881
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5918..6022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6023..6091,6092..6883)
                     /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(6241..6260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7054..7642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7543..7562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      241 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
      301 gaacaaaagc tggagctctt gcatggaggc gcaattggca aagacgtaaa gctcttgtca
      361 tcaattagat ctaatttgga aacagattgg tacaatttgg atggaatttt aggcttggca
      421 cctatcaatt gtgagcgacg ctttcttaca ttcaataact gttggttctg attttcactt
      481 tgactattgc cttggtatgg agaatttctc tgcaatggtt ttgcctgctt cgaaggagtg
      541 ctgttactat cgaagataaa ttccttcttt tcaaattgcc attgtaacat ggagttattc
      601 tctatatcct ggacgactgg aggagtcaag ggcctggaaa gtacggatcc tgtagaaata
      661 tcactggcaa ttatactgag tttattgttg gtgttgctat tcatatttga gctggtattg
      721 gacgaaagca aggataatga gtttgtgtct actgactcca acaccctagg ctggatttga
      781 tctttctcca tcctttaatt ttaacctttt aattagtggt tggatcaagt tttcgagact
      841 attccaatct gtgacttgtt ggataaatag tttttgactc gtttagtata atcctttttt
      901 ctaaaagtgc ttagagttct ctaagatgtt cttgtttaca atgtgagcga tttaggaaat
      961 ttcctaaaaa tggccgaggc ggcgctagca ttctacgaaa ggtgagataa cgcttcgtta
     1021 tcgaaaaatg tcaggggaca ggggttatat aagaacgaaa attgtcatcc tgcatttttt
     1081 ctttaaaaca gctatacaaa aagtgatacc gacatacaat gaagtatcta aacttagttt
     1141 tcgtgcttca gcttcttatt agcatcaaat acgcctcatt cggccgagcc ttttctcttt
     1201 ttgaagatga taccaccttt gccaatttgg ataaacagct aaagcttcca cagaatacac
     1261 agcaaaccct taaattggac cgtttgaatc acgatgatcc gctgtttaca acttttattt
     1321 cttctgtgga cacagattac agtttgagac ttagaacagt agatccttct aaactaggaa
     1381 ttgacaccgt aaaacaatgg tcgggttaca tggactataa ggattccaaa cacttttttt
     1441 actggttttt tgaaagtagg aacgatcctg ctaacgaccc aattattctt tggttaaatg
     1501 gtggacctgg ttgttcctcg tttactgggt tgctatttga actaggcccc tcatcaattg
     1561 gcgccgatat gaaaccaatc cacaatccct attcttggaa taatcaggct tcaatgatct
     1621 tcttagaaca gccactcgga gtcggctttt cctatggtga tgaaaaagtc tcctctacaa
     1681 aattagcagg caaagatgcg tacattttcc tggaattgtt ttttgaagct tttcctcatt
     1741 tacgctccaa cgatttccac attgcaggcg aatcctatgc aggacattat atccctcaaa
     1801 ttgcacatga gatcgttgtc aagaaccctg aaagaacgtt ccagttaact tcagttatga
     1861 ttggtaatgg tatcacagac cctttgattc aagcagatta ttatgaacca atggcatgcg
     1921 ggaaaggggg ctatcaccct gttctctcat cagaagaatg tgagaaaatg agtaaagctg
     1981 caggtcgttg tcgtaggttg aacaagttat gttatgcttc taaatcaagt ttaccatgca
     2041 tagtcgccac tgcttactgt gactctgcac ttttggaacc gtacattaac acaggactca
     2101 acgtctatga cattagaggg ccctgtgaag ataatagtac tgatggtatg tgttatacag
     2161 gtctccgcta tgtcgaccag tatatgaatt ttcctgaagt tcaagaaacg ctagggtccg
     2221 acgtgcataa ttattctggc tgtgataatg acgtgttcac cggatttttg tttacgggcg
     2281 atggaagtaa accatttcaa caatatattg ctgaattatt aaatcacaac attccggtat
     2341 taatatatgc gggtgataag gattatattt gtaattggct gggaaaccat gcttggtcca
     2401 atgagttgga atggatcaat aaacgtaggt atcagagaag gatgttaaga ccatgggtca
     2461 gtaaagaaac aggtgaagag ttgggacaag tcaagaacta tggccctttc acctttttga
     2521 gaatatacga tgccggtcat atggtgccct atgatcaacc ggaggcaagt ttggaaatgg
     2581 tcaacagttg gatttccggt aatcgtgctt tttcggatct ttccaccttg gaaaatgcta
     2641 gttctagacg gatccccggg ttaattaagc ttgcgcaaca cgatgaagcc gtggacaaca
     2701 aattcaacaa agaacaacaa aacgcgttct atgagatctt acatttacct aacttaaacg
     2761 aagaacaacg aaacgccttc atccaaagct taaaagatga cccaagccaa agcgctaacc
     2821 ttttagcaga agctaaaaag ctaaatgatg ctcaggcgcc gaaagtagac aacaaattca
     2881 acaaagaaca acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac
     2941 aacgaaacgc cgtcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag
     3001 cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacgcgaat taaggcgcgc
     3061 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     3121 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     3181 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct
     3241 tattgaccac acctctaccg gcagaattcg atatcaagct tatcgatacc gtcgacctcg
     3301 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     3361 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3421 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3481 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     3541 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3601 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3661 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3721 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3781 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3841 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3901 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3961 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg
     4021 gtaataactg atataattaa attgaagctc taatttgtga gtttagtata catgcattta
     4081 cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg
     4141 cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc
     4201 caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac
     4261 ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac
     4321 cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc
     4381 tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg
     4441 acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct
     4501 tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt
     4561 ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa
     4621 attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa
     4681 ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt
     4741 tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag
     4801 cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag
     4861 gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc
     4921 aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac
     4981 tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt
     5041 cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa
     5101 aaaatgatga attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc
     5161 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5221 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5281 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5341 ataggttaat gtcatgataa taatggtttc ttagacggat cgcttgcctg taacttacac
     5401 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
     5461 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
     5521 gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat
     5581 ttattagaca agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt
     5641 aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt
     5701 aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag
     5761 tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta
     5821 tttttaattt atatatttat attaaaaaat ttaaattata attattttta tagcacgtga
     5881 tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5941 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6001 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6061 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6121 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6181 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6241 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6301 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6361 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6421 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6481 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6541 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6601 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6661 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6721 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6781 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6841 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6901 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6961 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7201 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7381 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7441 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7501 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7561 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7621 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7681 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7741 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cga
//
