LOCUS       sequence-369819-        8579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ATG9-D830A/E831A mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8579)
  AUTHORS   Ren X, Nguyen TN, Lam WK, Buffalo CZ, Lazarou M, Yokom AL, Hurley JH
  TITLE     Structural basis for ATG9A recruitment to the ULK1 complex in 
            mitophagy initiation.
  JOURNAL   Sci Adv. 2023 Feb 15;9(7):eadg2997. doi: 10.1126/sciadv.adg2997. 
            Epub 2023 Feb 15.
  PUBMED    36791199
REFERENCE   2  (bases 1 to 8579)
  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"; journalName: "Sci Adv."; 
            date: "2023-02-15"; volume: "9(7):eadg2997. doi"; pages: " 
            10.1126/sciadv.adg2997. Epub 2023 Feb 15"
COMMENT     Sequence Label: pCAG-MBP-ATG9-D830A/E831A
FEATURES             Location/Qualifiers
     source          1..8579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1751..1760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1757..2857
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MGIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2831..2854
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2921..2941
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5600..5619)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5665..5720
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5719..5738)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6081..6097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6081..6097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6094..6116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6105..6121
                     /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(6129..6135,6136..6153,6154..6159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6174..6195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6254..6449
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6300..6435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6455..6589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6492..6511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6546..6565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6657..6674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6828..7416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6908..6927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7587..8378,8379..8447))
                     /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     8210..8229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8448..8552)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg ccrggggggg gggggsgggg sgrggggsgg
      541 ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgatcga
     1741 cggtatcgat gccaccatgg ggatcgaaga aggtaaactg gtaatctgga ttaacggcga
     1801 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
     1861 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
     1921 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
     1981 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
     2041 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
     2101 gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
     2161 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
     2221 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
     2281 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
     2341 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
     2401 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
     2461 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
     2521 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
     2581 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
     2641 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     2701 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     2761 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     2821 tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa
     2881 caataacaat aacaacaacc tcggggatat cccaacgacc gaaaacctgt attttcaggg
     2941 cgccatggga tccggtacca tggctcagtt cgacaccgaa taccagagat tggaagcgtc
     3001 atactccgac tcaccaccag gcgaagaaga tttgctcgtt catgttgctg agggttctaa
     3061 gtcaccttgg caccacatcg aaaacctgga cctgttcttc agccgcgtgt acaacctgca
     3121 ccagaagaac ggtttcacct gcatgctgat cggcgaaatc ttcgagctga tgcagttcct
     3181 gttcgtggtc gccttcacca ctttcctggt cagctgcgtg gactacgaca tcctgttcgc
     3241 taacaagatg gtgaaccact ctctgcaccc taccgagccc gtgaaggtca ctctgcccga
     3301 cgccttcctg ccagctcagg tgtgctccgc ccgcatccag gaaaacggct ctctgatcac
     3361 tatcctggtc atcgctggag tgttctggat tcacagactg atcaagttca tctacaacat
     3421 ctgctgctac tgggagatcc actcattcta cctgcacgcc ctgcgcatcc ctatgtccgc
     3481 tctgccctac tgcacctggc aggaggtgca ggctcgcatc gtccagactc agaaggaaca
     3541 ccaaatctgc atccacaagc gtgaactgac cgagctggac atctaccacc gcatcctgag
     3601 attccagaac tacatggtcg ccctggtgaa caagagcctg ctgccactga ggttcaggct
     3661 gcctggtctg ggtgaagctg tgttcttcac tcgtggtctg aagtacaact tcgaactgat
     3721 cctgttctgg ggacccggta gcctgttcct gaacgagtgg tccctgaagg ccgaatacaa
     3781 gaggggtggc cagagactgg agctggctca gcgcctgtct aaccgtatcc tgtggatcgg
     3841 catcgctaac ttcctgctgt gccccctgat cctgatctgg cagatcctgt atgccttctt
     3901 cagctacgct gaggtcctga agcgcgaacc aggagccctg ggtgctcgtt gctggtccct
     3961 gtacggaagg tgctacctga gacacttcaa cgagctggaa cacgagctgc aatcacgcct
     4021 gaaccgtggc tacaagccag cctcaaagta catgaactgc ttcctgtccc ctctgctgac
     4081 cctgctggct aagaacggcg ccttcttcgc tggttccatc ctggccgtgc tgatcgctct
     4141 gaccatctac gacgaggacg tcctggctgt ggaacacgtc ctgaccactg tcactctgct
     4201 gggcgtgacc gtcactgtgt gccgctcatt catccccgac cagcacatgg tgttctgccc
     4261 agagcagctg ctgcgtgtca tcctggccca catccactac atgcccgacc actggcaggg
     4321 aaacgctcac cgctcacaga cccgtgacga gttcgcccag ctgttccagt acaaggctgt
     4381 gttcatcctg gaggaactgc tgtcccctat cgtcactccc ctgatcctga tcttctgcct
     4441 gaggccaaga gccctggaga tcatcgactt cttccgcaac ttcaccgtgg aagtggtcgg
     4501 cgtcggagac acttgctctt tcgctcagat ggacgtgcgt cagcacggac acccacagtg
     4561 gctgtcagcc ggtcagaccg aggcttccgt ctaccagcag gccgaggacg gcaagaccga
     4621 actgtcactg atgcacttcg ctatcactaa ccctggatgg caacctccca gggagtccac
     4681 cgccttcctg ggtttcctga aggaacaggt gcagagagac ggtgctgccg ctagcctggc
     4741 tcagggaggt ctgctgcctg agaacgctct gttcacttcc atccagtccc tgcaatccga
     4801 aagcgagccc ctgtctctga tcgccaacgt ggtcgctggt tcctcctgcc gcggcccacc
     4861 tctgcctcgc gacctgcaag gaagccgcca cagggctgag gtggcttctg ctctgcgctc
     4921 tttctcccct ctgcaacctg gacaggctcc aactggtagg gctcactcaa ccatgactgg
     4981 ttccggcgtg gacgccagaa ctgcttcttc aggctccagc gtgtgggaag gacagctgca
     5041 atccctggtc ctgtccgagt acgccagcac cgaaatgtct ctgcacgctc tgtacatgca
     5101 ccagctgcac aagcagcagg cccaggctga accagagagg cacgtgtggc accgccgtga
     5161 atctgacgag tcaggagaat ctgctcctga cgagggcgga gaaggtgccc gcgctccaca
     5221 gtccatccct cgttccgcta gctacccatg cgctgctccc agaccaggcg cccccgaaac
     5281 cactgctctg cacggtggct tccagaggag atacggaggt atcaccgacc ccggaactgt
     5341 gcccagggtc ccaagccact tctcccgcct gccactgggc ggatgggctg aagacggaca
     5401 gtctgcttca cgtcaccctg aacccgtccc tgaagaaggt agcgaagccg cgttgccgcc
     5461 acaagtccat aaagtgtgag gtacccccgg gtgatatctt aattaattaa ctcgaggaat
     5521 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     5581 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     5641 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5701 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5761 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     5821 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     5881 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     5941 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     6001 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     6061 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6121 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6181 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6241 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6361 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6421 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     6481 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6541 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     6601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7261 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8521 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgg
//
