LOCUS       sequence-237869-        7933 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 7933)
  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 7933)
  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: pGAD-Atg17
FEATURES             Location/Qualifiers
     source          1..7933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..445
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             461..802
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     RBS             1873..1884
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2484..2671
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2539..2563)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2792..3886)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVRNPTHLNGIIITSNMFGDIVSDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3887..4292)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4334..4350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4334..4350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4347..4369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4358..4374
                     /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(4382..4388,4389..4406,4407..4412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4427..4448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4565..4582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4736..5324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4816..4835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5495..6286,6287..6355))
                     /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     6118..6137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6356..6460)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6528..6546
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6584..6606)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6706..6725
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6742..7906
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(7911..7933,1..379)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 tttcttttct ctctcccccg ttgttgtctc accatatccg caatgacaaa aaaatgatgg
       61 aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg ttgttccaga
      121 gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc acgcacacta
      181 ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg aaataaaaaa
      241 aagtttgctg tcttgctatc aagtataaat agacctgcaa ttattaatct tttgtttcct
      301 cgtcattgtt ctcgttccct ttcttccttg tttctttttc tgcacaatat ttcaagctat
      361 accaagcata caatcaactc caagctttgc aaagatggat aaagcggaat taattcccga
      421 gcctccaaaa aagaagagaa aggtcgaatt gggtaccgcc gccaatttta atcaaagtgg
      481 gaatattgct gatagctcat tgtccttcac tttcactaac agtagcaacg gtccgaacct
      541 cataacaact caaacaaatt ctcaagcgct ttcacaacca attgcctcct ctaacgttca
      601 tgataacttc atgaataatg aaatcacggc tagtaaaatt gatgatggta ataattcaaa
      661 accactgtca cctggttgga cggaccaaac tgcgtataac gcgtttggaa tcactacagg
      721 gatgtttaat accactacaa tggatgatgt atataactat ctattcgatg atgaagatac
      781 cccaccaaac ccaaaaaaag agatcgaatt ccccggggga tccatcgata acgaagcaga
      841 tgttacaaaa tttgttaata atgccaggaa aacgctgacc gatgctcaac ttttatgttc
      901 aagtgctaat ttaaggattg tagatattaa gaaaaaattg tcatcttggc agttgagtat
      961 ttcaaaactc aattttctaa tagttggctt gagacagcaa ggaaagtttc tttacactat
     1021 tttaaaggaa ggcattggga caaagctaat ccagaaacaa tggaatcaag ctgtcttagt
     1081 ggtcttagtt gatgagatga agtactggca atatgaaatc acttctaaag tacaaaggtt
     1141 ggatggtata gtaaatgaat tgagcatatc agaaaaagat gataccgatc cctctaaact
     1201 aggagattac atctcgagag acaacgttaa cttattgaat gacaaactga aagaagtgcc
     1261 agtaattgag cgtcaaattg aaaacattaa gcttcaatac gaaaatatgg tcagaaaagt
     1321 taacaaagaa ctgattgata ccaagttgac ggacgtaact caaaaattcc aaagtaaatt
     1381 tggtatagat aacctgatgg aaacaaatgt tgcagagcag tttagcaggg aactaacgga
     1441 ccttgaaaaa gatttagcag agataatgaa ttcattgaca cagcactttg ataagacatt
     1501 gctgttacaa gataaaaaaa ttgataacga tgaacgtgag gagctgttta aggtggtaca
     1561 aggcgacgac aaagaactat acaacatttt caaaactctg catgaggtaa ttgatgacgt
     1621 ggacaaaaca attcttaact tgggtcaatt tttgcaggca aaaataaagg aaaagacaga
     1681 actacacagc gaagtttctg aaataataaa cgatttcaat agaaatttgg aatatctatt
     1741 aatctttaaa gatatttcca atctgattga tagcttcaaa aattcctgta cacaagatat
     1801 tcaaacaact aaggaacttt gtgaatttta tgataatttt gaagaaagct acggtaactt
     1861 agttctagaa gcaaagagga gaaaggatgt ggcaaacaga atgaaaacta tattgaaaga
     1921 ttgtgaaaag cagttgcaga atttagatgc tcaggaccag gaagaacgtc agaattttat
     1981 agcggaaaat ggaacttatc ttcctgagac aatctggccc ggtaaaattg acgatttttc
     2041 ttccctgtac actttaaatt acaacgtgaa gaatccttag aaaaaaggaa aaaaaggctg
     2101 cagagatcta tgaatcgtag atactgaaaa accccgcaag ttcacttcaa ctgtgcatcg
     2161 tgcaccatct caatttcttt catttataca tcgttttgcc ttcttttatg taactatact
     2221 cctctaagtt tcaatcttgg ccatgtaacc tctgatctat agaatttttt aaatgactag
     2281 aattaatgcc catctttttt ttggacctaa attcttcatg aaaatatatt acgagggctt
     2341 attcagaagc tttggacttc ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg
     2401 gcttgtctac cttgccagaa atttacgaaa agatggaaaa gggtcaaatc gttggtagat
     2461 acgttgttga cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     2521 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     2581 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     2641 aggtcgctct tattgaccac acctctaccg gcatgccggt cgaaattccc ctaccctatg
     2701 aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta
     2761 tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta acctcttcgg
     2821 cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac
     2881 ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca
     2941 atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg accttattct
     3001 ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt
     3061 ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg
     3121 cttcatcgga gacgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg
     3181 ttgggttcct aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg
     3241 taggaaattc gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca
     3301 aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga
     3361 aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag
     3421 cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc actaattctc
     3481 tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa
     3541 gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga
     3601 tttttagtaa accttgttca ggtctaacac tacctgtacc ccatttagga ccacccacag
     3661 cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggga
     3721 cacctgtagc ttcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat
     3781 tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac
     3841 caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc
     3901 cttgaaatat atatatatat tgctgaaatg taaaaggtaa gaaaagttag aaagtaagac
     3961 gattgctaac cacctattgg aaaaaacaat aggtccttaa ataatattgt caacttcaag
     4021 tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc
     4081 ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg
     4141 cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc
     4201 cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat tcgaactctt
     4261 gcatcttacg atacctgagt attcccacag ttgggggatc tcgactctag ctagaggatc
     4321 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4381 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4441 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4501 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4561 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4621 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5161 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6481 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6541 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6601 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6661 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6721 ttgtactgag agtgcaccat aacgcattta agcataaaca cgcactatgc cgttcttctc
     6781 atgtatatat atatacaggc aacacgcaga tataggtgcg acgtgaacag tgagctgtat
     6841 gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt tgaagttcct
     6901 attccgaagt tcctattctc tagctagaaa gtataggaac ttcagagcgc ttttgaaaac
     6961 caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac gcaccggact gtaacgagct
     7021 actaaaatat tgcgaatacc gcttccacaa acattgctca aaagtatctc tttgctatat
     7081 atctctgtgc tatatcccta tataacctac ccatccacct ttcgctcctt gaacttgcat
     7141 ctaaactcga cctctacatt ttttatgttt atctctagta ttactcttta gacaaaaaaa
     7201 ttgtagtaag aactattcat agagtgaatc gaaaacaata cgaaaatgta aacatttcct
     7261 atacgtagta tatagagaca aaatagaaga aaccgttcat aattttctga ccaatgaaga
     7321 atcatcaacg ctatcacttt ctgttcacaa agtatgcgca atccacatcg gtatagaata
     7381 taatcgggga tgcctttatc ttgaaaaaat gcacccgcag cttcgctagt aatcagtaaa
     7441 cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa tagacaccaa agtagccttc
     7501 ttctaacctt aacggaccta cagtgcaaaa agttatcaag agactgcatt atagagcgca
     7561 caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa aatagcgctc tcgggatgca
     7621 tttttgtaga acaaaaaaga agtatagatt ctttgttggt aaaatagcgc tctcgcgttg
     7681 catttctgtt ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg
     7741 ttgcattttt gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg
     7801 cgttgcattt ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc
     7861 tcgcgttgca tttttgttct acaaaatgaa gcacagatgc ttcgttgctt gcatgcaact
     7921 tcttttcttt ttt
//
