LOCUS       sequence-238189-        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
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1945..1956
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2556..2743
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2611..2635)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2755..2966
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2967..3770
                     /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(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4844..5635,5636..5704))
                     /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     5467..5486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5705..5809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5877..5895
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5933..5955)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6055..6074
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6091..7255
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
      121 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
      181 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
      241 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct 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 taacgaagca gatgttacaa aatttgttaa taatgccagg aaaacgctga ccgatgctca
      961 acttttatgt tcaagtgcta atttaaggat tgtagatatt aagaaaaaat tgtcatcttg
     1021 gcagttgagt atttcaaaac tcaattttct aatagttggc ttgagacagc aaggaaagtt
     1081 tctttacact attttaaagg aaggcattgg gacaaagcta atccagaaac aatggaatca
     1141 agctgtctta gtggtcttag ttgatgagat gaagtactgg caatatgaaa tcacttctaa
     1201 agtacaaagg ttggatggta tagtaaatga attgagcata tcagaaaaag atgataccga
     1261 tccctctaaa ctaggagatt acatctcgag agacaacgtt aacttattga atgacaaact
     1321 gaaagaagtg ccagtaattg agcgtcaaat tgaaaacatt aagcttcaat acgaaaatat
     1381 ggtcagaaaa gttaacaaag aactgattga taccaagttg acggacgtaa ctcaaaaatt
     1441 ccaaagtaaa tttggtatag ataacctgat ggaaacaaat gttgcagagc agtttagcag
     1501 ggaactaacg gaccttgaaa aagatttagc agagataatg aattcattga cacagcactt
     1561 tgataagaca ttgctgttac aagataaaaa aattgataac gatgaacgtg aggagctgtt
     1621 taaggtggta caaggcgacg acaaagaact atacaacatt ttcaaaactc tgcatgaggt
     1681 aattgatgac gtggacaaaa caattcttaa cttgggtcaa tttttgcagg caaaaataaa
     1741 ggaaaagaca gaactacaca gcgaagtttc tgaaataata aacgatttca atagaaattt
     1801 ggaatatcta ttaatcttta aagatatttc caatctgatt gatagcttca aaaattcctg
     1861 tacacaagat attcaaacaa ctaaggaact ttgtgaattt tatgataatt ttgaagaaag
     1921 ctacggtaac ttagttctag aagcaaagag gagaaaggat gtggcaaaca gaatgaaaac
     1981 tatattgaaa gattgtgaaa agcagttgca gaatttagat gctcaggacc aggaagaacg
     2041 tcagaatttt atagcggaaa atggaactta tcttcctgag acaatctggc ccggtaaaat
     2101 tgacgatttt tcttccctgt acactttaaa ttacaacgtg aagaatcctt agaaaaaagg
     2161 aaaaaaaggc tgcagagatc tatgaatcgt agatactgaa aaaccccgca agttcacttc
     2221 aactgtgcat cgtgcaccat ctcaatttct ttcatttata catcgttttg ccttctttta
     2281 tgtaactata ctcctctaag tttcaatctt ggccatgtaa cctctgatct atagaatttt
     2341 ttaaatgact agaattaatg cccatctttt ttttggacct aaattcttca tgaaaatata
     2401 ttacgagggc ttattcagaa gctttggact tcttcgccag aggtttggtc aagtctccaa
     2461 tcaaggttgt cggcttgtct accttgccag aaatttacga aaagatggaa aagggtcaaa
     2521 tcgttggtag atacgttgtt gacacttcta aataagcgaa tttcttatga tttatgattt
     2581 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     2641 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
     2701 aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgcca gcttttcaat
     2761 tcatcatttt ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg
     2821 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     2881 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     2941 acaaaaaccg gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     3001 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     3061 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     3121 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     3181 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     3241 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     3301 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     3361 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     3421 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     3481 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3541 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     3601 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     3661 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3721 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     3781 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     3841 tattacccct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5821 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5881 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5941 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6001 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     6061 cagagcagat tgtactgaga gtgcaccata acgcatttaa gcataaacac gcactatgcc
     6121 gttcttctca tgtatatata tatacaggca acacgcagat ataggtgcga cgtgaacagt
     6181 gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt
     6241 gaagttccta ttccgaagtt cctattctct agctagaaag tataggaact tcagagcgct
     6301 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     6361 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     6421 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     6481 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     6541 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     6601 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     6661 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     6721 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     6781 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     6841 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     6901 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     6961 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     7021 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     7081 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     7141 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     7201 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgtt
//
