LOCUS       sequence-369646-        5419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To make ATG101/ATG13 HORMA complex for expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  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 5419)
  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-ATG13 (1-197)
FEATURES             Location/Qualifiers
     source          1..5419
                     /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             2283..2294
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2440..2459)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2505..2560
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2559..2578)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2921..2937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2934..2956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2945..2961
                     /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(2969..2975,2976..2993,2994..2999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3014..3035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3094..3289
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3140..3275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3295..3429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3332..3351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3386..3405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3497..3514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3668..4256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3748..3767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4427..5218,5219..5287))
                     /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     5050..5069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5288..5392)
                     /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 ccrggsgggg sggggsgggg sgrggggggg
      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 gggccgggga 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 agaccgacct caacagccag gaccgtaaag acctcgacaa
     1801 gttcattaag tttttcgccc tcaagaccgt gcaggtgatc gtgcaggccc ggctgggaga
     1861 gaaaatctgc acacgcagct cctcttcacc caccggttct gactggttca acctggctat
     1921 taaggatatc cctgaagtga ctcacgaagc aaagaaagca ctcgctggac agctgccagc
     1981 agtgggtcgt tcaatgtgtg tcgagatctc cctgaagaca tctgaaggag actcaatgga
     2041 gctcgaaatt tggtgcctgg agatgaacga aaagtgtgat aaagagatca aagtgagcta
     2101 caccgtctat aatcgtctgt ccctgctcct gaaatctctc ctggccatca ctcgggtgac
     2161 acctgcttac cgcctcagta ggaagcaggg tcatgaatat gtgattctgt acaggatcta
     2221 tttcggcgag gtccagctct ccggactggg tgaaggcttt cagactgtgc gtgtcggaac
     2281 cgtgggaact ccagtcggca caattaccct ctcttgcgca taccgtatca atctggcctt
     2341 catgtcatga ctcgaggaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     2401 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     2461 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     2521 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2581 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     2641 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     2701 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     2761 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     2821 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     2881 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3061 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     3121 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3181 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3241 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     3301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3421 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3481 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3541 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3601 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3661 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3721 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3781 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3841 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3901 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3961 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4021 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4081 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4141 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4201 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4261 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4321 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4381 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4441 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4501 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4561 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4621 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4681 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4741 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4801 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4861 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4921 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4981 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5041 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5101 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5161 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5221 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5281 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5341 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5401 cccgaaaagt gccacctgg
//
