LOCUS       sequence-231055-        7245 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For Yeast-two-hybrid analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7245)
  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 7245)
  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(F317D)
FEATURES             Location/Qualifiers
     source          1..7245
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             434..874
                     /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             1948..1959
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2542..2729
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2597..2621)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2740..2956
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2957..3760
                     /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(3904..3921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4075..4663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4155..4174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4834..5625,5626..5694))
                     /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     5457..5476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5695..5799)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5867..5885
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5923..5945)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6045..6064
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6081..7245
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
      241 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttccccgggg gatccatcga
      901 tatgaacgaa gcagatgtta caaaatttgt taataatgcc aggaaaacgc tgaccgatgc
      961 tcaactttta tgttcaagtg ctaatttaag gattgtagat attaagaaaa aattgtcatc
     1021 ttggcagttg agtatttcaa aactcaattt tctaatagtt ggcttgagac agcaaggaaa
     1081 gtttctttac actattttaa aggaaggcat tgggacaaag ctaatccaga aacaatggaa
     1141 tcaagctgtc ttagtggtct tagttgatga gatgaagtac tggcaatatg aaatcacttc
     1201 taaagtacaa aggttggatg gtatagtaaa tgaattgagc atatcagaaa aagatgatac
     1261 cgatccctct aaactaggag attacatctc gagagacaac gttaacttat tgaatgacaa
     1321 actgaaagaa gtgccagtaa ttgagcgtca aattgaaaac attaagcttc aatacgaaaa
     1381 tatggtcaga aaagttaaca aagaactgat tgataccaag ttgacggacg taactcaaaa
     1441 attccaaagt aaatttggta tagataacct gatggaaaca aatgttgcag agcagtttag
     1501 cagggaacta acggaccttg aaaaagattt agcagagata atgaattcat tgacacagca
     1561 ctttgataag acattgctgt tacaagataa aaaaattgat aacgatgaac gtgaggagct
     1621 gtttaaggtg gtacaaggcg acgacaaaga actatacaac attttcaaaa ctctgcatga
     1681 ggtaattgat gacgtggaca aaacaattct taacttgggt caatttttgc aggcaaaaat
     1741 aaaggaaaag acagaactac acagcgaagt ttctgaaata ataaacgatt tcaatagaaa
     1801 tttggaatat ctattaatct ttaaagatat ttccaatctg attgatagcg acaaaaattc
     1861 ctgtacacaa gatattcaaa caactaagga actttgtgaa ttttatgata attttgaaga
     1921 aagctacggt aacttagttc tagaagcaaa gaggagaaag gatgtggcaa acagaatgaa
     1981 aactatattg aaagattgtg aaaagcagtt gcagaattta gatgctcagg accaggaaga
     2041 acgtcagaat tttatagcgg aaaatggaac ttatcttcct gagacaatct ggcccggtaa
     2101 aattgacgat ttttcttccc tgtacacttt aaattacaac gtgaagaatc cttagctgca
     2161 gagatctatg aatcgtagat actgaaaaac cccgcaagtt cacttcaact gtgcatcgtg
     2221 caccatctca atttctttca tttatacatc gttttgcctt cttttatgta actatactcc
     2281 tctaagtttc aatcttggcc atgtaacctc tgatctatag aattttttaa atgactagaa
     2341 ttaatgccca tctttttttt ggacctaaat tcttcatgaa aatatattac gagggcttat
     2401 tcagaagctt tggacttctt cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc
     2461 ttgtctacct tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac
     2521 gttgttgaca cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa
     2581 gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa
     2641 ttcttgttct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag
     2701 gtcgctctta ttgaccacac ctctaccggc atgcagcttt tcaattcaat tcatcatttt
     2761 ttttttattc ttttttttga tttcggtttc tttgaaattt ttttgattcg gtaatctccg
     2821 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     2881 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaaccg
     2941 gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag
     3001 tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
     3061 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
     3121 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
     3181 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc
     3241 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     3301 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     3361 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     3421 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     3481 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     3541 cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca
     3601 acagtataga accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag
     3661 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     3721 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     3781 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccct
     3841 ggattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4501 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5821 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5881 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5941 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6001 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6061 tgtactgaga gtgcaccata acgcatttaa gcataaacac gcactatgcc gttcttctca
     6121 tgtatatata tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg
     6181 tgcgcagctc gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta
     6241 ttccgaagtt cctattctct agctagaaag tataggaact tcagagcgct tttgaaaacc
     6301 aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg taacgagcta
     6361 ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct ttgctatata
     6421 tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg aacttgcatc
     6481 taaactcgac ctctacattt tttatgttta tctctagtat tactctttag acaaaaaaat
     6541 tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa acatttccta
     6601 tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac caatgaagaa
     6661 tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat
     6721 aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac
     6781 gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa gtagccttct
     6841 tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta tagagcgcac
     6901 aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct cgggatgcat
     6961 ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct ctcgcgttgc
     7021 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     7081 tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata gcgctttcgc
     7141 gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct
     7201 cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgtt
//
