LOCUS       sequence-240332-        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(D195A,E196A)(405)
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     119..136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     120..136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     146..165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        146..164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2425..2598
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2599..2772
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      2808..2973
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2863..2887)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3036..3056)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3036..3054)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3075..3091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3075..3091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3088..3110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3099..3115
                     /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(3123..3129,3130..3147,3148..3153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3168..3189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3306..3323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3477..4065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3557..3576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4236..5027,5028..5096))
                     /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     4859..4878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5097..5201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5269..5287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5325..5347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5447..5466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5960..7054)
                     /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(7055..7462)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7763..8218)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7900..7921)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8112..8131
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
       61 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      121 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      181 gccccccctc gaggagattg tccctggtga gtctctgtct tttcctgcaa ttggccatta
      241 ttgcccacat ctgggtgcag gttaagattg agccctttca agttccttct ctgtaaagtc
      301 tttcgttgaa acatgaccaa caaccaatat accacgctgc aaaaacagat atgatccgat
      361 gcagttttct tatcattagt gtcgcttcac attaacctgt aggtcttcta atccatcaga
      421 tgggccaata atagtatgat atcgccgtta ccccacgaat cacttataac gaggacagtg
      481 atcaaaaaag ggagggaaaa aaatttcgta agaaaaagga caacacaatc ttagaactaa
      541 tgggcgattg taacattggg gtttgcaaga aaagacaatc atcaataaaa gtgtttatta
      601 tctgattgta catttatttc tcatcattgg attcagtttt gtttcatcca cattctttta
      661 agaccacaag cttaatattt tcattcataa aggattgtac gacaaaacaa caataatagt
      721 aaatatagtc atattttatt cctaatagaa agatagacgc agaaaattct tacttttagc
      781 ccttttattt cttcataacc gcatcgccat tcgcataata acttatccca ggcctaaaat
      841 acgacaagaa agaaaatagt agtaatgggt tccatcaata ataacttcaa cactaataat
      901 aacagtaata cggatctcga tcgggactgg aaaactgcat tgaatattcc caagaaggat
      961 accagaccac agacagatga tgtcttaaat acaaagggta atacttttga agatttttat
     1021 ttgaaaagag agctgttaat gggtattttt gaagccggtt ttgaaaaacc atcccctatt
     1081 caagaagagg ctattccggt agcaataact ggtagagaca tattggccag agcaaagaat
     1141 ggtacaggta agacagccgc atttgttatt cctacgttag agaaagtcaa accaaagttg
     1201 aataaaattc aggctttgat catggttccc acaagagagt tagctctgca aacttcccaa
     1261 gtcgtccgta cattgggaaa acactgtggt atttcatgta tggtgaccac tggtggtacc
     1321 aatttgagag acgatattct aagattaaat gaaacagtcc atattttggt tggtactcct
     1381 ggtagagtgt tggatttagc ttcgagaaaa gtagcagatc tgtctgattg ttcattattc
     1441 atcatggctg ctgccgataa aatgttatct cgtgacttca agacaataat tgaacaaatc
     1501 ttatccttct tacctccaac tcaccaatct ttattattta gcgctacttt cccactaacg
     1561 gttaaggaat tcatggttaa acatctgcat aagccttatg aaattaattt gatggaagaa
     1621 ttgactctaa agggtattac tcaatactac gcctttgttg aagaaagaca aaagctacat
     1681 tgtttaaata ctttattctc taagcttcaa attaatcaag ccattatttt ttgtaattct
     1741 actaaccgtg tcgaattact agccaagaaa atcactgatt taggctattc ctgttattac
     1801 tctcatgcaa gaatgaagca acaagaaaga aataaagttt tccacgaatt tcgtcaaggt
     1861 aaggttcgta cattggtctg ctccgattta ttgacccgtg gtatcgatat tcaagccgtt
     1921 aatgtcgtta tcaatttcga tttccccaaa acggcagaaa catatttaca tcgtattggt
     1981 agatccggta ggtttggcca cttgggttta gcaatcaatt tgattaactg gaatgatcgg
     2041 tttaaccttt ataaaatcga acaagaactg ggcactgaaa ttgcggctat tccggccact
     2101 atcgataagt cattgtatgt ggctgaaaat gacgaaacgg tacctgtccc attcccgata
     2161 gagcaacaga gctatcatca acaagcgata cctcagcaac aattaccatc tcagcaacaa
     2221 tttgctattc ctccgcagca acatcatcca caattcatgg ttcctccttc gcatcagcaa
     2281 caacaggcgt atcctccacc gcaaatgcca tcgcagcagg ggtatcctcc acagcaggaa
     2341 catttcatgg cgatgccacc tggtcagtca caaccccagt atcggatccc cgggttaatt
     2401 aagcttgcgc aacacgatga agccgtggac aacaaattca acaaagaaca acaaaacgcg
     2461 ttctatgaga tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa
     2521 agcttaaaag atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat
     2581 gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat
     2641 gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccgtcat ccaaagttta
     2701 aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgct
     2761 caggcgccga aagtagacgc gaattaaggc gcgccacttc taaataagcg aatttcttat
     2821 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     2881 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     2941 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacaccact agttctagag
     3001 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     3061 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3121 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3181 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3241 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3301 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3361 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5161 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5221 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5281 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5341 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5401 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5461 attgtactga gagtgcacca tatcgactac gtcgttaagg ccgtttctga cagagtaaaa
     5521 ttcttgaggg aactttcacc attatgggaa atggttcaag aaggtattga cttaaactcc
     5581 atcaaatggt caggtcattg agtgtttttt atttgttgta tttttttttt tttagagaaa
     5641 atcctccaat atataaatta ggaatcatag tttcatgatt ttctgttaca cctaactttt
     5701 tgtgtggtgc cctcctcctt gtcaatatta atgttaaagt gcaattcttt ttccttatca
     5761 cgttgagcca ttagtatcaa tttgcttacc tgtattcctt tacatcctcc tttttctcct
     5821 tcttgataaa tgtatgtaga ttgcgtatat agtttcgtct accctatgaa catattccat
     5881 tttgtaattt cgtgtcgttt ctattatgaa tttcatttat aaagtttatg tacaaatatc
     5941 ataaaaaaag agaatctttt taagcaagga ttttcttaac ttcttcggcg acagcatcac
     6001 cgacttcggt ggtactgttg gaaccaccta aatcaccagt tctgatacct gcatccaaaa
     6061 cctttttaac tgcatcttca atggccttac cttcttcagg caagttcaat gacaatttca
     6121 acatcattgc agcagacaag atagtggcga tagggttgac cttattcttt ggcaaatctg
     6181 gagcagaacc gtggcatggt tcgtacaaac caaatgcggt gttcttgtct ggcaaagagg
     6241 ccaaggacgc agatggcaac aaacccaagg aacctgggat aacggaggct tcatcggaga
     6301 tgatatcacc aaacatgttg ctggtgatta taataccatt taggtgggtt gggttcttaa
     6361 ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac cttcaatgta gggaattcgt
     6421 tcttgatggt ttcctccaca gtttttctcc ataatcttga agaggccaaa acattagctt
     6481 tatccaagga ccaaataggc aatggtggct catgttgtag ggccatgaaa gcggccattc
     6541 ttgtgattct ttgcacttct ggaacggtgt attgttcact atcccaagcg acaccatcac
     6601 catcgtcttc ctttctctta ccaaagtaaa tacctcccac taattctctg acaacaacga
     6661 agtcagtacc tttagcaaat tgtggcttga ttggagataa gtctaaaaga gagtcggatg
     6721 caaagttaca tggtcttaag ttggcgtaca attgaagttc tttacggatt tttagtaaac
     6781 cttgttcagg tctaacacta ccggtacccc atttaggacc acccacagca cctaacaaaa
     6841 cggcatcagc cttcttggag gcttccagcg cctcatctgg aagtggaaca cctgtagcat
     6901 cgatagcagc accaccaatt aaatgatttt cgaaatcgaa cttgacattg gaacgaacat
     6961 cagaaatagc tttaagaacc ttaatggctt cggctgtgat ttcttgacca acgtggtcac
     7021 ctggcaaaac gacgatcttc ttaggggcag acattagaat ggtatatcct tgaaatatat
     7081 atatatatat tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac gattgctaac
     7141 cacctattgg aaaaaacaat aggtccttaa ataatattgt caacttcaag tattgtgatg
     7201 caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc ggcgctctca
     7261 cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg cgacctctga
     7321 aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc cctgtgtgtt
     7381 ctcgttatgt tgaggaaaaa aataatggtt gctaagagat tcgaactctt gcatcttacg
     7441 atacctgagt attcccacag ttaactgcgg tcaagatatt tcttgaatca ggcgccttag
     7501 accgctcggc caaacaacca attacttgtt gagaaataga gtataattat cctataaata
     7561 taacgttttt gaacacacat gaacaaggaa gtacaggaca attgattttg aagagaatgt
     7621 ggattttgat gtaattgttg ggattccatt tttaataagg caataatatt aggtatgtag
     7681 atatactaga agttctcctc gaccggtcga tatgcggtgt gaaataccgc acagatgcgt
     7741 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     7801 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7861 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7921 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7981 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     8041 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     8101 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     8161 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     8221 attcgccatt caggctgcg
//
