LOCUS       sequence-369805-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To make monomeric ATG9 C-terminal tail for expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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 (692-839)
FEATURES             Location/Qualifiers
     source          1..6506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     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(3527..3546)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3592..3647
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3646..3665)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4008..4024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4008..4024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4021..4043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4032..4048
                     /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(4056..4062,4063..4080,4081..4086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4101..4122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4181..4376
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4227..4362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4382..4516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4419..4438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4473..4492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4584..4601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4755..5343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4835..4854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5514..6305,6306..6374))
                     /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     6137..6156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6375..6479)
                     /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 sggggsgggg sgrggggsgg
      541 ggsggggsga 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 tccggtaccc agctgcaatc cctggtcctg tccgagtacg ccagcaccga
     3001 aatgtctctg cacgctctgt acatgcacca gctgcacaag cagcaggccc aggctgaacc
     3061 agagaggcac gtgtggcacc gccgtgaatc tgacgagtca ggagaatctg ctcctgacga
     3121 gggcggagaa ggtgcccgcg ctccacagtc catccctcgt tccgctagct acccatgcgc
     3181 tgctcccaga ccaggcgccc ccgaaaccac tgctctgcac ggtggcttcc agaggagata
     3241 cggaggtatc accgaccccg gaactgtgcc cagggtccca agccacttct cccgcctgcc
     3301 actgggcgga tgggctgaag acggacagtc tgcttcacgt caccctgaac ccgtccctga
     3361 agaaggtagc gaagatgagt tgccgccaca agtccataaa gtgtgaggta cccccgggtg
     3421 atatcttaat taattaactc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg
     3481 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3541 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3601 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3661 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3721 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3781 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3841 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3901 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3961 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     4021 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4081 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4141 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4381 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4501 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4561 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4621 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4681 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4741 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4801 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4861 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4921 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4981 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5041 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5101 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5161 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5221 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5281 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5341 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5401 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5461 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5521 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5581 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5641 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5701 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5761 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5821 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5881 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5941 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6001 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6061 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6121 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6181 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6241 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6301 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6361 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6421 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6481 acatttcccc gaaaagtgcc acctgg
//
