LOCUS       sequence-400886-        6112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATG13 C-terminal and ULK1 MIT domain fusion 
            complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  AUTHORS   Chen M, Ren X, Nguyen TN, Khuu G, Cook ASI, Lazarou M, Hurley JH
  TITLE     Structure and activation of the human autophagy-initiating 
            ULK1C:PI3KC3-C1 supercomplex
  JOURNAL   bioRxiv 2023.06.01.543278
REFERENCE   2  (bases 1 to 6112)
  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: "bioRxiv 
            2023.06.01.543278"
COMMENT     Sequence Label: pCAG-ATG13 (363-517)-ULK1 (836-1050)-TSF
FEATURES             Location/Qualifiers
     source          1..6112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2891..2911
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2924..2947
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2984..3007
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3014..3037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3130..3149)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3195..3250
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3249..3268)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3611..3627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3611..3627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3624..3646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3635..3651
                     /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(3659..3665,3666..3683,3684..3689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3704..3725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3784..3979
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3830..3965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3985..4119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4022..4041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4076..4095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4187..4204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4358..4946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4438..4457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5117..5908,5909..5977))
                     /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     5740..5759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5978..6082)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgn caggcggggc ggggcggggc gaggggcggg
      541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc ctcgatcgac
     1741 ggtatcgata gccaccatgc acgatgtcct ggaaaccatc ttcgtgagaa aagtcggcgc
     1801 ttttgtgaac aagcccatta atcaggtcac actcacctct ctggacatcc ctttcgctat
     1861 gtttgcacca aagaacctcg agctggaaga cactgatccc atggtgaatc ctccagactc
     1921 acccgagaca gaaagtcctc tccagggcag tctgcacagc gatggaagct ccggtggctc
     1981 ttcaggcaac acacatgacg atttcgtcat gattgacttc aaacccgctt tttccaagga
     2041 cgatatcctc cctatggatc tgggaacctt ctacagggag tttcagaatc cccctcagct
     2101 cagtagcctg agcattgaca tcggtgccca gtccatggct gaggacctcg attctctgcc
     2161 tgaaaaactg gctgtgcacg agaagaacgt cagagaattc gacgcatttg tggagaccct
     2221 gcagggtagc gacgaagcga tggagcagga acacactgag atcctgaggg gtctcagatt
     2281 caccctgctc tttgtgcagc atgtcctgga aattgctgca ctcaagggta gtgctagcga
     2341 ggcagctggt ggaccagaat accagctgca ggagagcgtg gtcgccgacc agatctctct
     2401 gctctcaagg gaatggggct tcgctgagca gctggtcctc tatctgaagg tggccgaact
     2461 gctcagtagc ggcctgcagt ccgccatcga tcagattaga gctggaaagc tctgcctgtc
     2521 ctctactgtg aaacaggtgg tccggcgcct gaacgagctc tacaaggcca gtgtggtcag
     2581 ctgtcaggga ctctctctgc gtctccagcg gttctttctg gacaaacagc gcctgctcga
     2641 taggatccac tcaattacag ctgaacgcct gatcttcagc catgcagtgc agatggtcca
     2701 gtccgcagcc ctggacgaaa tgtttcagca cagggaggga tgcgtcccca gataccataa
     2761 ggctctgctc ctgctcgagg gtctgcagca catgctcagc gaccaggccg atatcgaaaa
     2821 tgtgactaag tgcaaactgt gtattgagag gagactctcc gctctgctca caggcatctg
     2881 tgcaggtacc gaaaacctgt acttccaatc cggtatggcg gcctggtcac acccgcagtt
     2941 cgagaaaggt ggaggtgccc gaggtggatc gggaggtgga tcgtggagcc acccgcagtt
     3001 cgaaaaagga ttcgattaca aggatgacga tgacaagtag ctcgaggaat tcactcctca
     3061 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     3121 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     3181 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3241 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     3301 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     3361 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     3421 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     3481 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     3541 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     3601 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3661 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3721 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     3781 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3901 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3961 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     4021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4081 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     4141 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4201 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4261 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4321 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4381 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4441 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4501 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4561 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4621 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4681 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4741 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4801 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4861 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4921 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4981 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5041 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5101 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5161 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5221 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5281 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5341 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5401 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5461 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5521 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5581 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5641 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5701 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5761 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5821 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5881 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5941 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6001 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6061 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tc
//
