LOCUS       sequence-389380-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-ATG9A-N terminal 
            cytosolic domain fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  AUTHORS   Mattera R, Park SY, De Pace R, Guardia CM, Bonifacino JS
  TITLE     AP-4 mediates export of ATG9A from the trans-Golgi network to 
            promote autophagosome formation.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Dec 12;114(50):E10697-E10706. doi: 
            10.1073/pnas.1717327114. Epub 2017 Nov 27.
  PUBMED    29180427
REFERENCE   2  (bases 1 to 5714)
  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: 
            "10.1073/pnas.1717327114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-12-12- 12"; volume: "114"; issue: "50"; pages: 
            "E10697-E10706"
COMMENT     Sequence Label: pGBT9-ATG9A-N
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(5..109)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     177..195
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(233..255)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     355..374
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      391..1555
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1560..1961
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1989..2429
                     /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"
     terminator      2667..2854
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2722..2746)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2907..3581)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3582..3683)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3697..3713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3697..3713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3710..3732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3721..3737
                     /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(3745..3751,3752..3769,3770..3775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3790..3811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3928..3945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4099..4687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4179..4198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4858..5649,5650..5714,1..4))
                     /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     5481..5500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
       61 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      121 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
      181 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
      241 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
      301 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      361 cagagcagat tgtactgaga gtgcaccata acgcatttaa gcataaacac gcactatgcc
      421 gttcttctca tgtatatata tatacaggca acacgcagat ataggtgcga cgtgaacagt
      481 gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt
      541 gaagttccta ttccgaagtt cctattctct agctagaaag tataggaact tcagagcgct
      601 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
      661 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
      721 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
      781 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
      841 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
      901 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
      961 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     1021 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     1081 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     1141 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     1201 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     1261 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     1321 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     1381 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     1441 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     1501 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttgcttg
     1561 catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc tcaccatatc
     1621 cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca aagacagcac
     1681 caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg aaactttttc
     1741 cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct tccttccagt
     1801 tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa atagacctgc
     1861 aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct tgtttctttt
     1921 tctgcacaat atttcaagct ataccaagca tacaatcaac tccaagcttg aagcaagcct
     1981 cctgaaagat gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa
     2041 agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt
     2101 gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg
     2161 aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg
     2221 acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg
     2281 tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata
     2341 tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta
     2401 acaaaggtca aagacagttg actgtatcgc cggaattcat ggcgcagttt gacactgaat
     2461 accagcgcct agaggcctcc tatagtgatt cacccccagg ggaggaggac ctgttggtgc
     2521 acgtcgccga ggggagcaag tcaccttggc accatattga aaaccttgac ctcttcttct
     2581 ctcgagttta taatctgcac cagaagaatg gcttcacatg tatgctcatc ggggagtgag
     2641 tcgacctgca gccaagctaa ttccgggcga atttcttatg atttatgatt tttattatta
     2701 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     2761 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     2821 atgaggtcgc tcttattgac cacacctcta ccggcatgcc ggcaagtgca caaacaatac
     2881 ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt
     2941 ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg
     3001 ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa
     3061 tgtaagcttt cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa
     3121 agttcacctg tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt
     3181 gaagctgcac tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc
     3241 aaaccgagga actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca
     3301 atgccgtaat cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca
     3361 accaagtatt tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc
     3421 cttgcaataa ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag
     3481 tcagcatcgg aatctagagc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc
     3541 accaatggac cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg
     3601 tgcttaatca cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct
     3661 ccttttcttt tttcgaccga attaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     3721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4501 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5701 gaaatgttga atac
//
