LOCUS       sequence-238194-        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(L105D)
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             353..793
                     /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             1864..1875
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2475..2662
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2530..2554)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2674..2885
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2886..3689
                     /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(3833..3850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4004..4592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4084..4103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4763..5554,5555..5623))
                     /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     5386..5405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5624..5728)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5796..5814
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5852..5874)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5974..5993
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6010..7174
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(7179..7255,1..325)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt
       61 tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc
      121 acactactct ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat
      181 aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat taatcttttg
      241 tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca caatatttca
      301 agctatacca agcatacaat caactccaag cttgaagcaa gcctcctgaa agatgaagct
      361 actgtcttct atcgaacaag catgcgatat ttgccgactt aaaaagctca agtgctccaa
      421 agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct actctcccaa
      481 aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa ggctagaaag
      541 actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga ttttgaaaat
      601 ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag ataatgtgaa
      661 taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc taacattgag
      721 acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag gtcaaagaca
      781 gttgactgta tcgccggaat tccccggggg atccatcgat aacgaagcag atgttacaaa
      841 atttgttaat aatgccagga aaacgctgac cgatgctcaa cttttatgtt caagtgctaa
      901 tttaaggatt gtagatatta agaaaaaatt gtcatcttgg cagttgagta tttcaaaact
      961 caattttcta atagttggct tgagacagca aggaaagttt ctttacacta ttttaaagga
     1021 aggcattggg acaaagctaa tccagaaaca atggaatcaa gctgtcttag tggtcttagt
     1081 tgatgagatg aagtactggc aatatgaaat cacttctaaa gtacaaaggg acgatggtat
     1141 agtaaatgaa ttgagcatat cagaaaaaga tgataccgat ccctctaaac taggagatta
     1201 catctcgaga gacaacgtta acttattgaa tgacaaactg aaagaagtgc cagtaattga
     1261 gcgtcaaatt gaaaacatta agcttcaata cgaaaatatg gtcagaaaag ttaacaaaga
     1321 actgattgat accaagttga cggacgtaac tcaaaaattc caaagtaaat ttggtataga
     1381 taacctgatg gaaacaaatg ttgcagagca gtttagcagg gaactaacgg accttgaaaa
     1441 agatttagca gagataatga attcattgac acagcacttt gataagacat tgctgttaca
     1501 agataaaaaa attgataacg atgaacgtga ggagctgttt aaggtggtac aaggcgacga
     1561 caaagaacta tacaacattt tcaaaactct gcatgaggta attgatgacg tggacaaaac
     1621 aattcttaac ttgggtcaat ttttgcaggc aaaaataaag gaaaagacag aactacacag
     1681 cgaagtttct gaaataataa acgatttcaa tagaaatttg gaatatctat taatctttaa
     1741 agatatttcc aatctgattg atagcttcaa aaattcctgt acacaagata ttcaaacaac
     1801 taaggaactt tgtgaatttt atgataattt tgaagaaagc tacggtaact tagttctaga
     1861 agcaaagagg agaaaggatg tggcaaacag aatgaaaact atattgaaag attgtgaaaa
     1921 gcagttgcag aatttagatg ctcaggacca ggaagaacgt cagaatttta tagcggaaaa
     1981 tggaacttat cttcctgaga caatctggcc cggtaaaatt gacgattttt cttccctgta
     2041 cactttaaat tacaacgtga agaatcctta gaaaaaagga aaaaaaggct gcagagatct
     2101 atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc gtgcaccatc
     2161 tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac tcctctaagt
     2221 ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta gaattaatgc
     2281 ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct tattcagaag
     2341 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2401 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2461 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2521 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2581 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2641 ttattgacca cacctctacc ggcatgccag cttttcaatt catcattttt tttttattct
     2701 tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     2761 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     2821 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaaccgg aaacgaagat
     2881 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     2941 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     3001 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     3061 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     3121 cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta
     3181 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     3241 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     3301 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     3361 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     3421 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     3481 ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa
     3541 ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg
     3601 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     3661 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     3721 taaactcaca aattagagct tcaatttaat tatatcagtt attacccctg cattaatgaa
     3781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5761 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5821 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5941 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6001 tgcaccataa cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
     6061 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
     6121 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
     6181 ctattctcta gctagaaagt ataggaactt cagagcgctt ttgaaaacca aaagcgctct
     6241 gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg
     6301 cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta
     6361 tatccctata taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc
     6421 tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa
     6481 ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata
     6541 tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct
     6601 atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg
     6661 cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg
     6721 agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa
     6781 cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa
     6841 aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac
     6901 aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct
     6961 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     7021 tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct
     7081 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     7141 tttgttctac aaaatgaagc acagatgctt cgttgcttgc atgcaacttc ttttcttttt
     7201 ttttcttttc tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaa
//
