LOCUS       sequence-238188-        7255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Yeast-two-hybrid analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7255)
  AUTHORS   Liu X, Mao K, Yu AYH, Omairi-Nasser A, Austin J 2nd, Glick BS, Yip 
            CK, Klionsky DJ
  TITLE     The Atg17-Atg31-Atg29 Complex Coordinates with Atg11 to Recruit the 
            Vam7 SNARE and Mediate Autophagosome-Vacuole Fusion.
  JOURNAL   Curr Biol. 2016 Jan 25;26(2):150-160. doi: 
            10.1016/j.cub.2015.11.054. Epub 2016 Jan 7.
  PUBMED    26774783
REFERENCE   2  (bases 1 to 7255)
  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"; doi: 
            "10.1016/j.cub.2015.11.054"; journalName: "Curr Biol"; date: 
            "2016-01-25- 25"; volume: "26"; issue: "2"; pages: "150-160"
COMMENT     Sequence Label: pGBDU-Atg17(I325D)
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..411
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             439..879
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     RBS             1950..1961
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2561..2748
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2616..2640)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2760..2971
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2972..3775
                     /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"
     primer_bind     complement(3919..3936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4090..4678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4170..4189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4849..5640,5641..5709))
                     /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     5472..5491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5710..5814)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5882..5900
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5938..5960)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6060..6079
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(6096..7255,1..5)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgttgcttg catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc
       61 tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca
      121 aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg
      181 aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct
      241 tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa
      301 atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct
      361 tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac tccaagcttg
      421 aagcaagcct cctgaaagat gaagctactg tcttctatcg aacaagcatg cgatatttgc
      481 cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac
      541 aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg
      601 acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga
      661 gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca
      721 ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg
      781 gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa
      841 gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc cgggggatcc
      901 atcgataacg aagcagatgt tacaaaattt gttaataatg ccaggaaaac gctgaccgat
      961 gctcaacttt tatgttcaag tgctaattta aggattgtag atattaagaa aaaattgtca
     1021 tcttggcagt tgagtatttc aaaactcaat tttctaatag ttggcttgag acagcaagga
     1081 aagtttcttt acactatttt aaaggaaggc attgggacaa agctaatcca gaaacaatgg
     1141 aatcaagctg tcttagtggt cttagttgat gagatgaagt actggcaata tgaaatcact
     1201 tctaaagtac aaaggttgga tggtatagta aatgaattga gcatatcaga aaaagatgat
     1261 accgatccct ctaaactagg agattacatc tcgagagaca acgttaactt attgaatgac
     1321 aaactgaaag aagtgccagt aattgagcgt caaattgaaa acattaagct tcaatacgaa
     1381 aatatggtca gaaaagttaa caaagaactg attgatacca agttgacgga cgtaactcaa
     1441 aaattccaaa gtaaatttgg tatagataac ctgatggaaa caaatgttgc agagcagttt
     1501 agcagggaac taacggacct tgaaaaagat ttagcagaga taatgaattc attgacacag
     1561 cactttgata agacattgct gttacaagat aaaaaaattg ataacgatga acgtgaggag
     1621 ctgtttaagg tggtacaagg cgacgacaaa gaactataca acattttcaa aactctgcat
     1681 gaggtaattg atgacgtgga caaaacaatt cttaacttgg gtcaattttt gcaggcaaaa
     1741 ataaaggaaa agacagaact acacagcgaa gtttctgaaa taataaacga tttcaataga
     1801 aatttggaat atctattaat ctttaaagat atttccaatc tgattgatag cttcaaaaat
     1861 tcctgtacac aagatgatca aacaactaag gaactttgtg aattttatga taattttgaa
     1921 gaaagctacg gtaacttagt tctagaagca aagaggagaa aggatgtggc aaacagaatg
     1981 aaaactatat tgaaagattg tgaaaagcag ttgcagaatt tagatgctca ggaccaggaa
     2041 gaacgtcaga attttatagc ggaaaatgga acttatcttc ctgagacaat ctggcccggt
     2101 aaaattgacg atttttcttc cctgtacact ttaaattaca acgtgaagaa tccttagaaa
     2161 aaaggaaaaa aaggctgcag agatctatga atcgtagata ctgaaaaacc ccgcaagttc
     2221 acttcaactg tgcatcgtgc accatctcaa tttctttcat ttatacatcg ttttgccttc
     2281 ttttatgtaa ctatactcct ctaagtttca atcttggcca tgtaacctct gatctataga
     2341 attttttaaa tgactagaat taatgcccat cttttttttg gacctaaatt cttcatgaaa
     2401 atatattacg agggcttatt cagaagcttt ggacttcttc gccagaggtt tggtcaagtc
     2461 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     2521 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     2581 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     2641 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     2701 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccagcttt
     2761 tcaattcatc attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg
     2821 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     2881 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     2941 acagaacaaa aaccggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
     3001 tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
     3061 aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
     3121 attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
     3181 ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
     3241 agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
     3301 cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
     3361 tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
     3421 agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
     3481 cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
     3541 aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
     3601 agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat
     3661 tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
     3721 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
     3781 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
     3841 tcagttatta cccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5881 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5941 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6061 ggcatcagag cagattgtac tgagagtgca ccataacgca tttaagcata aacacgcact
     6121 atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg tgcgacgtga
     6181 acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac
     6241 gctttgaagt tcctattccg aagttcctat tctctagcta gaaagtatag gaacttcaga
     6301 gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg
     6361 gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta
     6421 tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct
     6481 ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc
     6541 tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa
     6601 tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt
     6661 ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac
     6721 atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc
     6781 tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca
     6841 ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg
     6901 cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc
     6961 gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata
     7021 gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     7081 ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta
     7141 aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg
     7201 aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgct
//
