LOCUS       sequence-201594-        7353 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Prc1 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 7353)
  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 7353)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pKP136 (pRS406-PRC1p-PRC1-PA-ADH1t)
FEATURES             Location/Qualifiers
     source          1..7353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             4724..4897
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             4898..5071
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      5107..5294
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5162..5186)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5317..5333)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5318..5334)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5358..5377)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5359..5377)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5387..5404)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5387..5403)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5396..5418)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5548..6003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5635..5654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5845..5866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6134..6937)
                     /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(6938..7158)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7184..7203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7303..7325
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcg
     2281 aatgagtgaa aaatttgtta gcagagaagc aagaaggaaa gaaggaaaaa aaaaagaaac
     2341 cgtgtaagaa atcatggcgc ctcaaaaaaa aaaaaatacg cgaaatcgat ttccgtatat
     2401 gatgatacat atgttaggtc tcttatggta gtttttaggg tctgtcctgt tttttgaagg
     2461 cattggttag ggtctagtaa gtaactcgta taaagagatg tacttttata cggtatgtgt
     2521 ggcggttatt tccagtgttt tttttccctt attttgtggg ttcatggaaa tagtacatat
     2581 tgaccttctg cacaagaagc catattgaca gagcagtatg tgaggacctt tcttctacac
     2641 aatgtaacag tcaatggttg tagtcaatac cacgaccttt acgtgcagtt ttagagcgaa
     2701 acttcggttt tttgagactt acctcgtgtt tgtgtctccc tgggaatgga gccgcgccac
     2761 ccttgtctat tatgtatagt cttatttaat catagcgatg ttggtcatcc agtacacttc
     2821 ggtagcaacc ttcgtttgtg attgtcttgg taattgcttc caacaacttt atccatcatt
     2881 gagacagggg ccatatcacc cgcggggtct caaagaaggg gcccactaat aaaagcacga
     2941 gataagaatg ccagcaaaaa agctccgaaa taattctttt cgtcttccct cctagtctta
     3001 acaagacaag agagagagag agaaagaata ctcactagag attgtttctt ttctactcaa
     3061 cttaaagtat acatacgcta tgaaagcatt caccagttta ctatgtggac taggcctgtc
     3121 cactacactc gctaaggcca tctcattgca aagaccgttg ggtctagata aggacgtttt
     3181 gctgcaagct gcggaaaaat ttggtttgga cctcgacctg gatcatctct tgaaggagtt
     3241 ggactccaat gtattggacg cttgggccca aatagagcat ttgtacccaa accaggttat
     3301 gagccttgaa acttccacta agccaaaatt ccctgaagca atcaaaacga agaaagactg
     3361 ggactttgtg gtcaagaatg acgcaattga aaactatcag cttcgtgtca acaagattaa
     3421 ggaccctaaa atcctgggca ttgacccaaa tgtcacacag tacacgggtt acttggatgt
     3481 ggaagacgag gacaagcatt tcttcttttg gacttttgaa agtagaaacg atcctgcaaa
     3541 ggatccggtc atcctttggt tgaacggggg tccaggttgt tcttcactaa ccgggctgtt
     3601 ctttgaatta ggaccctcat ccattggacc tgatttgaaa cccatcggga acccttactc
     3661 ttggaacagc aatgccaccg tgatcttcct tgaccagcct gtcaacgttg ggttctcgta
     3721 ttccgggtcc tcaggtgttt ccaacactgt cgccgctggt aaggatgtct ataacttctt
     3781 ggagttgttc ttcgatcagt tccctgaata cgtcaacaag ggccaagatt tccacatcgc
     3841 tggggaatcc tacgccggcc attacatccc tgtttttgcc tctgaaattt tgtctcacaa
     3901 ggacagaaac ttcaacttaa cctccgtctt gatcggaaat ggcctcactg acccattgac
     3961 tcagtataac tattacgaac caatggcctg tggtgaaggt ggcgaaccat ctgttttgcc
     4021 ctcggaggaa tgctctgcta tggaagactc tttggaacgt tgtttgggct tgatcgagtc
     4081 gtgctatgac tcgcaatccg tctggtcctg tgttccagct accatttatt gtaataacgc
     4141 ccaattggct ccttaccaac gtaccggcag aaacgtttac gatatcagga aggattgtga
     4201 aggtggcaat ttgtgctacc caacgttaca agatatcgac gactacttaa accaggacta
     4261 cgtcaaagaa gctgtcggtg cggaggttga ccactacgaa tcctgtaact tcgatatcaa
     4321 cagaaatttc ctgtttgcgg gtgattggat gaagccttac cacaccgccg taacagatct
     4381 tttgaatcaa gacctaccca ttctggtata tgcaggcgat aaagatttca tctgtaactg
     4441 gttgggtaat aaggcgtgga cggatgtctt gccatggaag tacgacgaag aatttgcaag
     4501 ccaaaaagta cgtaactgga ctgcttctat caccgacgag gtcgctggtg aagtcaaatc
     4561 ctacaagcac ttcacctatt tgagagtctt caatggtggc cacatggttc catttgacgt
     4621 ccctgaaaac gccttaagta tggttaacga atggatccac ggtggtttct ccttatctag
     4681 acggatcccc gggttaatta agcttgcgca acacgatgaa gccgtggaca acaaattcaa
     4741 caaagaacaa caaaacgcgt tctatgagat cttacattta cctaacttaa acgaagaaca
     4801 acgaaacgcc ttcatccaaa gcttaaaaga tgacccaagc caaagcgcta accttttagc
     4861 agaagctaaa aagctaaatg atgctcaggc gccgaaagta gacaacaaat tcaacaaaga
     4921 acaacaaaac gcgttctatg agatcttaca tttacctaac ttaaacgaag aacaacgaaa
     4981 cgccgtcatc caaagtttaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc
     5041 taaaaagcta aatgatgctc aggcgccgaa agtagacgcg aattaaggcg cgccacttct
     5101 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     5161 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     5221 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     5281 cacacctcta ccggcagaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg
     5341 gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     5401 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5461 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5521 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
     5581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5941 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     6001 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta
     6061 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     6121 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     6181 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     6241 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     6301 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     6361 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     6421 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     6481 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     6541 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     6601 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     6661 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     6721 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     6781 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     6841 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     6901 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     6961 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     7021 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     7081 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
     7141 atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc tgctctgatg
     7201 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7261 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7321 agaggttttc accgtcatca ccgaaacgcg cga
//
