LOCUS       sequence-240368-        8239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Dhh1-PA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  AUTHORS   Liu X, Yao Z, Jin M, Namkoong S, Yin Z, Lee JH, Klionsky DJ
  TITLE     Dhh1 promotes autophagy-related protein translation during nitrogen 
            starvation.
  JOURNAL   PLoS Biol. 2019 Apr 11;17(4):e3000219. doi: 
            10.1371/journal.pbio.3000219. eCollection 2019 Apr.
  PUBMED    30973873
REFERENCE   2  (bases 1 to 8239)
  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: "PLoS 
            Biol."; date: "2019-04-11"; volume: "17(4):e3000219. doi"; pages: " 
            10.1371/journal.pbio.3000219. eCollection 2019 Apr"
COMMENT     Sequence Label: pRS-Dhh1-PA (405)
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     167..186
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        167..185
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2446..2619
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2620..2793
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      2829..2994
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2884..2908)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3057..3077)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3057..3075)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3096..3112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3096..3112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3109..3131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3120..3136
                     /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(3144..3150,3151..3168,3169..3174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3189..3210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3327..3344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3498..4086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3578..3597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4257..5048,5049..5117))
                     /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     4880..4899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5118..5222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5290..5308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5346..5368)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5468..5487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5981..7075)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(7076..7483)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7784..8239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7921..7942)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8133..8152
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
      181 atagggcgaa ttgggtaccg ggccccccct cgaggagatt gtccctggtg agtctctgtc
      241 ttttcctgca attggccatt attgcccaca tctgggtgca ggttaagatt gagccctttc
      301 aagttccttc tctgtaaagt ctttcgttga aacatgacca acaaccaata taccacgctg
      361 caaaaacaga tatgatccga tgcagttttc ttatcattag tgtcgcttca cattaacctg
      421 taggtcttct aatccatcag atgggccaat aatagtatga tatcgccgtt accccacgaa
      481 tcacttataa cgaggacagt gatcaaaaaa gggagggaaa aaaatttcgt aagaaaaagg
      541 acaacacaat cttagaacta atgggcgatt gtaacattgg ggtttgcaag aaaagacaat
      601 catcaataaa agtgtttatt atctgattgt acatttattt ctcatcattg gattcagttt
      661 tgtttcatcc acattctttt aagaccacaa gcttaatatt ttcattcata aaggattgta
      721 cgacaaaaca acaataatag taaatatagt catattttat tcctaataga aagatagacg
      781 cagaaaattc ttacttttag cccttttatt tcttcataac cgcatcgcca ttcgcataat
      841 aacttatccc aggcctaaaa tacgacaaga aagaaaatag tagtaatggg ttccatcaat
      901 aataacttca acactaataa taacagtaat acggatctcg atcgggactg gaaaactgca
      961 ttgaatattc ccaagaagga taccagacca cagacagatg atgtcttaaa tacaaagggt
     1021 aatacttttg aagattttta tttgaaaaga gagctgttaa tgggtatttt tgaagccggt
     1081 tttgaaaaac catcccctat tcaagaagag gctattccgg tagcaataac tggtagagac
     1141 atattggcca gagcaaagaa tggtacaggt aagacagccg catttgttat tcctacgtta
     1201 gagaaagtca aaccaaagtt gaataaaatt caggctttga tcatggttcc cacaagagag
     1261 ttagctctgc aaacttccca agtcgtccgt acattgggaa aacactgtgg tatttcatgt
     1321 atggtgacca ctggtggtac caatttgaga gacgatattc taagattaaa tgaaacagtc
     1381 catattttgg ttggtactcc tggtagagtg ttggatttag cttcgagaaa agtagcagat
     1441 ctgtctgatt gttcattatt catcatggac gaagccgata aaatgttatc tcgtgacttc
     1501 aagacaataa ttgaacaaat cttatccttc ttacctccaa ctcaccaatc tttattattt
     1561 agcgctactt tcccactaac ggttaaggaa ttcatggtta aacatctgca taagccttat
     1621 gaaattaatt tgatggaaga attgactcta aagggtatta ctcaatacta cgcctttgtt
     1681 gaagaaagac aaaagctaca ttgtttaaat actttattct ctaagcttca aattaatcaa
     1741 gccattattt tttgtaattc tactaaccgt gtcgaattac tagccaagaa aatcactgat
     1801 ttaggctatt cctgttatta ctctcatgca agaatgaagc aacaagaaag aaataaagtt
     1861 ttccacgaat ttcgtcaagg taaggttcgt acattggtct gctccgattt attgacccgt
     1921 ggtatcgata ttcaagccgt taatgtcgtt atcaatttcg atttccccaa aacggcagaa
     1981 acatatttac atcgtattgg tagatccggt aggtttggcc acttgggttt agcaatcaat
     2041 ttgattaact ggaatgatcg gtttaacctt tataaaatcg aacaagaact gggcactgaa
     2101 attgcggcta ttccggccac tatcgataag tcattgtatg tggctgaaaa tgacgaaacg
     2161 gtacctgtcc cattcccgat agagcaacag agctatcatc aacaagcgat acctcagcaa
     2221 caattaccat ctcagcaaca atttgctatt cctccgcagc aacatcatcc acaattcatg
     2281 gttcctcctt cgcatcagca acaacaggcg tatcctccac cgcaaatgcc atcgcagcag
     2341 gggtatcctc cacagcagga acatttcatg gcgatgccac ctggtcagtc acaaccccag
     2401 tatcggatcc ccgggttaat taagcttgcg caacacgatg aagccgtgga caacaaattc
     2461 aacaaagaac aacaaaacgc gttctatgag atcttacatt tacctaactt aaacgaagaa
     2521 caacgaaacg ccttcatcca aagcttaaaa gatgacccaa gccaaagcgc taacctttta
     2581 gcagaagcta aaaagctaaa tgatgctcag gcgccgaaag tagacaacaa attcaacaaa
     2641 gaacaacaaa acgcgttcta tgagatctta catttaccta acttaaacga agaacaacga
     2701 aacgccgtca tccaaagttt aaaagatgac ccaagccaaa gcgctaacct tttagcagaa
     2761 gctaaaaagc taaatgatgc tcaggcgccg aaagtagacg cgaattaagg cgcgccactt
     2821 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     2881 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     2941 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     3001 accacaccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     3061 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3661 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3901 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5221 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5281 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5341 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5401 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     5461 aactatgcgg catcagagca gattgtactg agagtgcacc atatcgacta cgtcgttaag
     5521 gccgtttctg acagagtaaa attcttgagg gaactttcac cattatggga aatggttcaa
     5581 gaaggtattg acttaaactc catcaaatgg tcaggtcatt gagtgttttt tatttgttgt
     5641 attttttttt ttttagagaa aatcctccaa tatataaatt aggaatcata gtttcatgat
     5701 tttctgttac acctaacttt ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag
     5761 tgcaattctt tttccttatc acgttgagcc attagtatca atttgcttac ctgtattcct
     5821 ttacatcctc ctttttctcc ttcttgataa atgtatgtag attgcgtata tagtttcgtc
     5881 taccctatga acatattcca ttttgtaatt tcgtgtcgtt tctattatga atttcattta
     5941 taaagtttat gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg attttcttaa
     6001 cttcttcggc gacagcatca ccgacttcgg tggtactgtt ggaaccacct aaatcaccag
     6061 ttctgatacc tgcatccaaa acctttttaa ctgcatcttc aatggcctta ccttcttcag
     6121 gcaagttcaa tgacaatttc aacatcattg cagcagacaa gatagtggcg atagggttga
     6181 ccttattctt tggcaaatct ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg
     6241 tgttcttgtc tggcaaagag gccaaggacg cagatggcaa caaacccaag gaacctggga
     6301 taacggaggc ttcatcggag atgatatcac caaacatgtt gctggtgatt ataataccat
     6361 ttaggtgggt tgggttctta actaggatca tggcggcaga atcaatcaat tgatgttgaa
     6421 ccttcaatgt agggaattcg ttcttgatgg tttcctccac agtttttctc cataatcttg
     6481 aagaggccaa aacattagct ttatccaagg accaaatagg caatggtggc tcatgttgta
     6541 gggccatgaa agcggccatt cttgtgattc tttgcacttc tggaacggtg tattgttcac
     6601 tatcccaagc gacaccatca ccatcgtctt cctttctctt accaaagtaa atacctccca
     6661 ctaattctct gacaacaacg aagtcagtac ctttagcaaa ttgtggcttg attggagata
     6721 agtctaaaag agagtcggat gcaaagttac atggtcttaa gttggcgtac aattgaagtt
     6781 ctttacggat ttttagtaaa ccttgttcag gtctaacact accggtaccc catttaggac
     6841 cacccacagc acctaacaaa acggcatcag ccttcttgga ggcttccagc gcctcatctg
     6901 gaagtggaac acctgtagca tcgatagcag caccaccaat taaatgattt tcgaaatcga
     6961 acttgacatt ggaacgaaca tcagaaatag ctttaagaac cttaatggct tcggctgtga
     7021 tttcttgacc aacgtggtca cctggcaaaa cgacgatctt cttaggggca gacattagaa
     7081 tggtatatcc ttgaaatata tatatatata ttgctgaaat gtaaaaggta agaaaagtta
     7141 gaaagtaaga cgattgctaa ccacctattg gaaaaaacaa taggtcctta aataatattg
     7201 tcaacttcaa gtattgtgat gcaagcattt agtcatgaac gcttctctat tctatatgaa
     7261 aagccggttc cggcgctctc acctttcctt tttctcccaa tttttcagtt gaaaaaggta
     7321 tatgcgtcag gcgacctctg aaattaacaa aaaatttcca gtcatcgaat ttgattctgt
     7381 gcgatagcgc ccctgtgtgt tctcgttatg ttgaggaaaa aaataatggt tgctaagaga
     7441 ttcgaactct tgcatcttac gatacctgag tattcccaca gttaactgcg gtcaagatat
     7501 ttcttgaatc aggcgcctta gaccgctcgg ccaaacaacc aattacttgt tgagaaatag
     7561 agtataatta tcctataaat ataacgtttt tgaacacaca tgaacaagga agtacaggac
     7621 aattgatttt gaagagaatg tggattttga tgtaattgtt gggattccat ttttaataag
     7681 gcaataatat taggtatgta gatatactag aagttctcct cgaccggtcg atatgcggtg
     7741 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     7801 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7861 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     7921 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7981 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     8041 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     8101 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     8161 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     8221 gcgccgctac agggcgcgt
//
