LOCUS       sequence-240331-        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(S226A,T228A)(405)
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     79..96
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     106..125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2385..2558
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2559..2732
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      2768..2933
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2823..2847)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2996..3016)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2996..3014)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3035..3051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3035..3051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3048..3070)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3059..3075
                     /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(3083..3089,3090..3107,3108..3113))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3128..3149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3266..3283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3437..4025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3517..3536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4196..4987,4988..5056))
                     /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     4819..4838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5057..5161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5229..5247
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5285..5307)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5407..5426
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5920..7014)
                     /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(7015..7422)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7723..8178)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7860..7881)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8072..8091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
      121 tagggcgaat tgggtaccgg gccccccctc gaggagattg tccctggtga gtctctgtct
      181 tttcctgcaa ttggccatta ttgcccacat ctgggtgcag gttaagattg agccctttca
      241 agttccttct ctgtaaagtc tttcgttgaa acatgaccaa caaccaatat accacgctgc
      301 aaaaacagat atgatccgat gcagttttct tatcattagt gtcgcttcac attaacctgt
      361 aggtcttcta atccatcaga tgggccaata atagtatgat atcgccgtta ccccacgaat
      421 cacttataac gaggacagtg atcaaaaaag ggagggaaaa aaatttcgta agaaaaagga
      481 caacacaatc ttagaactaa tgggcgattg taacattggg gtttgcaaga aaagacaatc
      541 atcaataaaa gtgtttatta tctgattgta catttatttc tcatcattgg attcagtttt
      601 gtttcatcca cattctttta agaccacaag cttaatattt tcattcataa aggattgtac
      661 gacaaaacaa caataatagt aaatatagtc atattttatt cctaatagaa agatagacgc
      721 agaaaattct tacttttagc ccttttattt cttcataacc gcatcgccat tcgcataata
      781 acttatccca ggcctaaaat acgacaagaa agaaaatagt agtaatgggt tccatcaata
      841 ataacttcaa cactaataat aacagtaata cggatctcga tcgggactgg aaaactgcat
      901 tgaatattcc caagaaggat accagaccac agacagatga tgtcttaaat acaaagggta
      961 atacttttga agatttttat ttgaaaagag agctgttaat gggtattttt gaagccggtt
     1021 ttgaaaaacc atcccctatt caagaagagg ctattccggt agcaataact ggtagagaca
     1081 tattggccag agcaaagaat ggtacaggta agacagccgc atttgttatt cctacgttag
     1141 agaaagtcaa accaaagttg aataaaattc aggctttgat catggttccc acaagagagt
     1201 tagctctgca aacttcccaa gtcgtccgta cattgggaaa acactgtggt atttcatgta
     1261 tggtgaccac tggtggtacc aatttgagag acgatattct aagattaaat gaaacagtcc
     1321 atattttggt tggtactcct ggtagagtgt tggatttagc ttcgagaaaa gtagcagatc
     1381 tgtctgattg ttcattattc atcatggacg aagccgataa aatgttatct cgtgacttca
     1441 agacaataat tgaacaaatc ttatccttct tacctccaac tcaccaatct ttattatttg
     1501 ctgctgcatt cccactaacg gttaaggaat tcatggttaa acatctgcat aagccttatg
     1561 aaattaattt gatggaagaa ttgactctaa agggtattac tcaatactac gcctttgttg
     1621 aagaaagaca aaagctacat tgtttaaata ctttattctc taagcttcaa attaatcaag
     1681 ccattatttt ttgtaattct actaaccgtg tcgaattact agccaagaaa atcactgatt
     1741 taggctattc ctgttattac tctcatgcaa gaatgaagca acaagaaaga aataaagttt
     1801 tccacgaatt tcgtcaaggt aaggttcgta cattggtctg ctccgattta ttgacccgtg
     1861 gtatcgatat tcaagccgtt aatgtcgtta tcaatttcga tttccccaaa acggcagaaa
     1921 catatttaca tcgtattggt agatccggta ggtttggcca cttgggttta gcaatcaatt
     1981 tgattaactg gaatgatcgg tttaaccttt ataaaatcga acaagaactg ggcactgaaa
     2041 ttgcggctat tccggccact atcgataagt cattgtatgt ggctgaaaat gacgaaacgg
     2101 tacctgtccc attcccgata gagcaacaga gctatcatca acaagcgata cctcagcaac
     2161 aattaccatc tcagcaacaa tttgctattc ctccgcagca acatcatcca caattcatgg
     2221 ttcctccttc gcatcagcaa caacaggcgt atcctccacc gcaaatgcca tcgcagcagg
     2281 ggtatcctcc acagcaggaa catttcatgg cgatgccacc tggtcagtca caaccccagt
     2341 atcggatccc cgggttaatt aagcttgcgc aacacgatga agccgtggac aacaaattca
     2401 acaaagaaca acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac
     2461 aacgaaacgc cttcatccaa agcttaaaag atgacccaag ccaaagcgct aaccttttag
     2521 cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag
     2581 aacaacaaaa cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa
     2641 acgccgtcat ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag
     2701 ctaaaaagct aaatgatgct caggcgccga aagtagacgc gaattaaggc gcgccacttc
     2761 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     2821 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     2881 taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga
     2941 ccacaccact agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     3001 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3061 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3121 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3181 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3241 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3421 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3841 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4201 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4261 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4321 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4381 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4441 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4501 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4561 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4621 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4681 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4741 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4801 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4861 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4921 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4981 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5041 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5161 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5221 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5281 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5341 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5401 actatgcggc atcagagcag attgtactga gagtgcacca tatcgactac gtcgttaagg
     5461 ccgtttctga cagagtaaaa ttcttgaggg aactttcacc attatgggaa atggttcaag
     5521 aaggtattga cttaaactcc atcaaatggt caggtcattg agtgtttttt atttgttgta
     5581 tttttttttt tttagagaaa atcctccaat atataaatta ggaatcatag tttcatgatt
     5641 ttctgttaca cctaactttt tgtgtggtgc cctcctcctt gtcaatatta atgttaaagt
     5701 gcaattcttt ttccttatca cgttgagcca ttagtatcaa tttgcttacc tgtattcctt
     5761 tacatcctcc tttttctcct tcttgataaa tgtatgtaga ttgcgtatat agtttcgtct
     5821 accctatgaa catattccat tttgtaattt cgtgtcgttt ctattatgaa tttcatttat
     5881 aaagtttatg tacaaatatc ataaaaaaag agaatctttt taagcaagga ttttcttaac
     5941 ttcttcggcg acagcatcac cgacttcggt ggtactgttg gaaccaccta aatcaccagt
     6001 tctgatacct gcatccaaaa cctttttaac tgcatcttca atggccttac cttcttcagg
     6061 caagttcaat gacaatttca acatcattgc agcagacaag atagtggcga tagggttgac
     6121 cttattcttt ggcaaatctg gagcagaacc gtggcatggt tcgtacaaac caaatgcggt
     6181 gttcttgtct ggcaaagagg ccaaggacgc agatggcaac aaacccaagg aacctgggat
     6241 aacggaggct tcatcggaga tgatatcacc aaacatgttg ctggtgatta taataccatt
     6301 taggtgggtt gggttcttaa ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac
     6361 cttcaatgta gggaattcgt tcttgatggt ttcctccaca gtttttctcc ataatcttga
     6421 agaggccaaa acattagctt tatccaagga ccaaataggc aatggtggct catgttgtag
     6481 ggccatgaaa gcggccattc ttgtgattct ttgcacttct ggaacggtgt attgttcact
     6541 atcccaagcg acaccatcac catcgtcttc ctttctctta ccaaagtaaa tacctcccac
     6601 taattctctg acaacaacga agtcagtacc tttagcaaat tgtggcttga ttggagataa
     6661 gtctaaaaga gagtcggatg caaagttaca tggtcttaag ttggcgtaca attgaagttc
     6721 tttacggatt tttagtaaac cttgttcagg tctaacacta ccggtacccc atttaggacc
     6781 acccacagca cctaacaaaa cggcatcagc cttcttggag gcttccagcg cctcatctgg
     6841 aagtggaaca cctgtagcat cgatagcagc accaccaatt aaatgatttt cgaaatcgaa
     6901 cttgacattg gaacgaacat cagaaatagc tttaagaacc ttaatggctt cggctgtgat
     6961 ttcttgacca acgtggtcac ctggcaaaac gacgatcttc ttaggggcag acattagaat
     7021 ggtatatcct tgaaatatat atatatatat tgctgaaatg taaaaggtaa gaaaagttag
     7081 aaagtaagac gattgctaac cacctattgg aaaaaacaat aggtccttaa ataatattgt
     7141 caacttcaag tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa
     7201 agccggttcc ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat
     7261 atgcgtcagg cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg
     7321 cgatagcgcc cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat
     7381 tcgaactctt gcatcttacg atacctgagt attcccacag ttaactgcgg tcaagatatt
     7441 tcttgaatca ggcgccttag accgctcggc caaacaacca attacttgtt gagaaataga
     7501 gtataattat cctataaata taacgttttt gaacacacat gaacaaggaa gtacaggaca
     7561 attgattttg aagagaatgt ggattttgat gtaattgttg ggattccatt tttaataagg
     7621 caataatatt aggtatgtag atatactaga agttctcctc gaccggtcga tatgcggtgt
     7681 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     7741 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     7801 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     7861 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     7921 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     7981 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     8041 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     8101 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     8161 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8221 ggtgcgggcc tcttcgcta
//
