LOCUS       sequence-231517-        7672 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 7672)
  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 7672)
  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(∆335-417)
FEATURES             Location/Qualifiers
     source          1..7672
                     /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             452..472
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             488..829
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     terminator      2254..2441
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2309..2333)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2559..3653)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAFEATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3654..4058)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4100..4116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4100..4116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4113..4135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4124..4140
                     /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(4148..4154,4155..4172,4173..4178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4331..4348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4502..5090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4582..4601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6294..6312
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6350..6372)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6472..6491
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6508..7672
                     /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 gctttgcaaa
      421 gatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg
      481 taccgccgcc aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt
      541 cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc
      601 acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag
      661 taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc
      721 gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata
      781 taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagaga tcgaattccc
      841 cgggggatcc atcgatatga acgaagcaga tgttacaaaa tttgttaata atgccaggaa
      901 aacgctgacc gatgctcaac ttttatgttc aagtgctaat ttaaggattg tagatattaa
      961 gaaaaaattg tcatcttggc agttgagtat ttcaaaactc aattttctaa tagttggctt
     1021 gagacagcaa ggaaagtttc tttacactat tttaaaggaa ggcattggga caaagctaat
     1081 ccagaaacaa tggaatcaag ctgtcttagt ggtcttagtt gatgagatga agtactggca
     1141 atatgaaatc acttctaaag tacaaaggtt ggatggtata gtaaatgaat tgagcatatc
     1201 agaaaaagat gataccgatc cctctaaact aggagattac atctcgagag acaacgttaa
     1261 cttattgaat gacaaactga aagaagtgcc agtaattgag cgtcaaattg aaaacattaa
     1321 gcttcaatac gaaaatatgg tcagaaaagt taacaaagaa ctgattgata ccaagttgac
     1381 ggacgtaact caaaaattcc aaagtaaatt tggtatagat aacctgatgg aaacaaatgt
     1441 tgcagagcag tttagcaggg aactaacgga ccttgaaaaa gatttagcag agataatgaa
     1501 ttcattgaca cagcactttg ataagacatt gctgttacaa gataaaaaaa ttgataacga
     1561 tgaacgtgag gagctgttta aggtggtaca aggcgacgac aaagaactat acaacatttt
     1621 caaaactctg catgaggtaa ttgatgacgt ggacaaaaca attcttaact tgggtcaatt
     1681 tttgcaggca aaaataaagg aaaagacaga actacacagc gaagtttctg aaataataaa
     1741 cgatttcaat agaaatttgg aatatctatt aatctttaaa gatatttcca atctgattga
     1801 tagcttcaaa aattcctgta cacaagatat tcaaacaact aaggaacttt gtgaatttta
     1861 ggtcgacctg cagagatcta tgaatcgtag atactgaaaa accccgcaag ttcacttcaa
     1921 ctgtgcatcg tgcaccatct caatttcttt catttataca tcgttttgcc ttcttttatg
     1981 taactatact cctctaagtt tcaatcttgg ccatgtaacc tctgatctat agaatttttt
     2041 aaatgactag aattaatgcc catctttttt ttggacctaa attcttcatg aaaatatatt
     2101 acgagggctt attcagaagc tttggacttc ttcgccagag gtttggtcaa gtctccaatc
     2161 aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa gggtcaaatc
     2221 gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt tatgattttt
     2281 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     2341 ttaaaacgaa aattcttgtt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     2401 gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgcccga aattccccta
     2461 ccctatgaac atattccatt ttgtaatttc gtgtcgtttc tattatgaat ttcatttata
     2521 aagtttatgt acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact
     2581 tcttcggcga cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt
     2641 ctgatacctg catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc
     2701 aagttcaatg acaatttcaa catcattgca gcagacaaga tagtggcgat agggtcaacc
     2761 ttattctttg gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg
     2821 ttcttgtctg gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata
     2881 acggaggctt catcggagat gatatcacca aacatgttgc tggtgattat aataccattt
     2941 aggtgggttg ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc
     3001 ttcaatgtag gaaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa
     3061 gaggccaaaa cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg
     3121 gccatgaaag cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta
     3181 tcccaagcga caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact
     3241 aattctctga caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag
     3301 tctaaaagag agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct
     3361 ttacggattt ttagtaaacc ttgttcaggt ctaacactac ctgtacccca tttaggacca
     3421 cccacagcac ctaacaaaac ggcatcaacc ttcttggagg cttccagcgc ctcatctgga
     3481 agtgggacac ctgtagcttc gaaagcagca ccaccaatta aatgattttc gaaatcgaac
     3541 ttgacattgg aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt
     3601 tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct taggggcaga cattagaatg
     3661 gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa
     3721 gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa
     3781 cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc
     3841 cggttccggc ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc
     3901 gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat
     3961 agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga
     4021 actcttgcat cttacgatac ctgagtattc ccacagttgg ggatctcgac tctagctaga
     4081 ggatcaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4201 ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4261 gccagctgga ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4921 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6241 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6301 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6361 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6421 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6481 agcagattgt actgagagtg caccataacg catttaagca taaacacgca ctatgccgtt
     6541 cttctcatgt atatatatat acaggcaaca cgcagatata ggtgcgacgt gaacagtgag
     6601 ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc gttttcggaa acgctttgaa
     6661 gttcctattc cgaagttcct attctctagc tagaaagtat aggaacttca gagcgctttt
     6721 gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa
     6781 cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg
     6841 ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac
     6901 ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca
     6961 aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca
     7021 tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa
     7081 tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat
     7141 agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc
     7201 agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta
     7261 gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag
     7321 agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg
     7381 gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa tagcgctctc
     7441 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     7501 ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg taaaatagcg
     7561 ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta
     7621 gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg tt
//
