LOCUS       sequence-240382-        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,S226A,T228A)(405)
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     115..132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     116..132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2421..2594
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2595..2768
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAVIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     terminator      2804..2969
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2859..2883)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3032..3052)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3032..3050)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3071..3087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3071..3087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3084..3106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3095..3111
                     /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(3119..3125,3126..3143,3144..3149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3164..3185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3302..3319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3473..4061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3553..3572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4232..5023,5024..5092))
                     /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     4855..4874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5093..5197)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5265..5283
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5321..5343)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5443..5462
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5956..7050)
                     /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(7051..7458)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7759..8214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7896..7917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8108..8127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
       61 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
      121 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
      181 cccctcgagg agattgtccc tggtgagtct ctgtcttttc ctgcaattgg ccattattgc
      241 ccacatctgg gtgcaggtta agattgagcc ctttcaagtt ccttctctgt aaagtctttc
      301 gttgaaacat gaccaacaac caatatacca cgctgcaaaa acagatatga tccgatgcag
      361 ttttcttatc attagtgtcg cttcacatta acctgtaggt cttctaatcc atcagatggg
      421 ccaataatag tatgatatcg ccgttacccc acgaatcact tataacgagg acagtgatca
      481 aaaaagggag ggaaaaaaat ttcgtaagaa aaaggacaac acaatcttag aactaatggg
      541 cgattgtaac attggggttt gcaagaaaag acaatcatca ataaaagtgt ttattatctg
      601 attgtacatt tatttctcat cattggattc agttttgttt catccacatt cttttaagac
      661 cacaagctta atattttcat tcataaagga ttgtacgaca aaacaacaat aatagtaaat
      721 atagtcatat tttattccta atagaaagat agacgcagaa aattcttact tttagccctt
      781 ttatttcttc ataaccgcat cgccattcgc ataataactt atcccaggcc taaaatacga
      841 caagaaagaa aatagtagta atgggttcca tcaataataa cttcaacact aataataaca
      901 gtaatacgga tctcgatcgg gactggaaaa ctgcattgaa tattcccaag aaggatacca
      961 gaccacagac agatgatgtc ttaaatacaa agggtaatac ttttgaagat ttttatttga
     1021 aaagagagct gttaatgggt atttttgaag ccggttttga aaaaccatcc cctattcaag
     1081 aagaggctat tccggtagca ataactggta gagacatatt ggccagagca aagaatggta
     1141 caggtaagac agccgcattt gttattccta cgttagagaa agtcaaacca aagttgaata
     1201 aaattcaggc tttgatcatg gttcccacaa gagagttagc tctgcaaact tcccaagtcg
     1261 tccgtacatt gggaaaacac tgtggtattt catgtatggt gaccactggt ggtaccaatt
     1321 tgagagacga tattctaaga ttaaatgaaa cagtccatat tttggttggt actcctggta
     1381 gagtgttgga tttagcttcg agaaaagtag cagatctgtc tgattgttca ttattcatca
     1441 tggctgctgc cgataaaatg ttatctcgtg acttcaagac aataattgaa caaatcttat
     1501 ccttcttacc tccaactcac caatctttat tatttgctgc tgcattccca ctaacggtta
     1561 aggaattcat ggttaaacat ctgcataagc cttatgaaat taatttgatg gaagaattga
     1621 ctctaaaggg tattactcaa tactacgcct ttgttgaaga aagacaaaag ctacattgtt
     1681 taaatacttt attctctaag cttcaaatta atcaagccat tattttttgt aattctacta
     1741 accgtgtcga attactagcc aagaaaatca ctgatttagg ctattcctgt tattactctc
     1801 atgcaagaat gaagcaacaa gaaagaaata aagttttcca cgaatttcgt caaggtaagg
     1861 ttcgtacatt ggtctgctcc gatttattga cccgtggtat cgatattcaa gccgttaatg
     1921 tcgttatcaa tttcgatttc cccaaaacgg cagaaacata tttacatcgt attggtagat
     1981 ccggtaggtt tggccacttg ggtttagcaa tcaatttgat taactggaat gatcggttta
     2041 acctttataa aatcgaacaa gaactgggca ctgaaattgc ggctattccg gccactatcg
     2101 ataagtcatt gtatgtggct gaaaatgacg aaacggtacc tgtcccattc ccgatagagc
     2161 aacagagcta tcatcaacaa gcgatacctc agcaacaatt accatctcag caacaatttg
     2221 ctattcctcc gcagcaacat catccacaat tcatggttcc tccttcgcat cagcaacaac
     2281 aggcgtatcc tccaccgcaa atgccatcgc agcaggggta tcctccacag caggaacatt
     2341 tcatggcgat gccacctggt cagtcacaac cccagtatcg gatccccggg ttaattaagc
     2401 ttgcgcaaca cgatgaagcc gtggacaaca aattcaacaa agaacaacaa aacgcgttct
     2461 atgagatctt acatttacct aacttaaacg aagaacaacg aaacgccttc atccaaagct
     2521 taaaagatga cccaagccaa agcgctaacc ttttagcaga agctaaaaag ctaaatgatg
     2581 ctcaggcgcc gaaagtagac aacaaattca acaaagaaca acaaaacgcg ttctatgaga
     2641 tcttacattt acctaactta aacgaagaac aacgaaacgc cgtcatccaa agtttaaaag
     2701 atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat gatgctcagg
     2761 cgccgaaagt agacgcgaat taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     2821 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2881 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2941 gctttctcag gtatagtatg aggtcgctct tattgaccac accactagtt ctagagcggc
     3001 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5221 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5281 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5341 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5401 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     5461 tactgagagt gcaccatatc gactacgtcg ttaaggccgt ttctgacaga gtaaaattct
     5521 tgagggaact ttcaccatta tgggaaatgg ttcaagaagg tattgactta aactccatca
     5581 aatggtcagg tcattgagtg ttttttattt gttgtatttt ttttttttta gagaaaatcc
     5641 tccaatatat aaattaggaa tcatagtttc atgattttct gttacaccta actttttgtg
     5701 tggtgccctc ctccttgtca atattaatgt taaagtgcaa ttctttttcc ttatcacgtt
     5761 gagccattag tatcaatttg cttacctgta ttcctttaca tcctcctttt tctccttctt
     5821 gataaatgta tgtagattgc gtatatagtt tcgtctaccc tatgaacata ttccattttg
     5881 taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca aatatcataa
     5941 aaaaagagaa tctttttaag caaggatttt cttaacttct tcggcgacag catcaccgac
     6001 ttcggtggta ctgttggaac cacctaaatc accagttctg atacctgcat ccaaaacctt
     6061 tttaactgca tcttcaatgg ccttaccttc ttcaggcaag ttcaatgaca atttcaacat
     6121 cattgcagca gacaagatag tggcgatagg gttgacctta ttctttggca aatctggagc
     6181 agaaccgtgg catggttcgt acaaaccaaa tgcggtgttc ttgtctggca aagaggccaa
     6241 ggacgcagat ggcaacaaac ccaaggaacc tgggataacg gaggcttcat cggagatgat
     6301 atcaccaaac atgttgctgg tgattataat accatttagg tgggttgggt tcttaactag
     6361 gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggga attcgttctt
     6421 gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat tagctttatc
     6481 caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg ccattcttgt
     6541 gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac catcaccatc
     6601 gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa caacgaagtc
     6661 agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt cggatgcaaa
     6721 gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta gtaaaccttg
     6781 ttcaggtcta acactaccgg taccccattt aggaccaccc acagcaccta acaaaacggc
     6841 atcagccttc ttggaggctt ccagcgcctc atctggaagt ggaacacctg tagcatcgat
     6901 agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac gaacatcaga
     6961 aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt ggtcacctgg
     7021 caaaacgacg atcttcttag gggcagacat tagaatggta tatccttgaa atatatatat
     7081 atatattgct gaaatgtaaa aggtaagaaa agttagaaag taagacgatt gctaaccacc
     7141 tattggaaaa aacaataggt ccttaaataa tattgtcaac ttcaagtatt gtgatgcaag
     7201 catttagtca tgaacgcttc tctattctat atgaaaagcc ggttccggcg ctctcacctt
     7261 tcctttttct cccaattttt cagttgaaaa aggtatatgc gtcaggcgac ctctgaaatt
     7321 aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat agcgcccctg tgtgttctcg
     7381 ttatgttgag gaaaaaaata atggttgcta agagattcga actcttgcat cttacgatac
     7441 ctgagtattc ccacagttaa ctgcggtcaa gatatttctt gaatcaggcg ccttagaccg
     7501 ctcggccaaa caaccaatta cttgttgaga aatagagtat aattatccta taaatataac
     7561 gtttttgaac acacatgaac aaggaagtac aggacaattg attttgaaga gaatgtggat
     7621 tttgatgtaa ttgttgggat tccattttta ataaggcaat aatattaggt atgtagatat
     7681 actagaagtt ctcctcgacc ggtcgatatg cggtgtgaaa taccgcacag atgcgtaagg
     7741 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     7801 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     7861 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     7921 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7981 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     8041 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     8101 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     8161 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     8221 gccattcagg ctgcgcaac
//
