LOCUS       sequence-436291-         783 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of TwinStrep-Flag -ATG13 C-terminal and ULK1 
            MIT domain fusion complex fused with GFP-10His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 783)
  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 783)
  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-TSF-ATG13 (363-517)-ULK1 (836-1050)-msfGFP
FEATURES             Location/Qualifiers
     source          1..783
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             7..27
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(34..36,37..39,40..750)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(79..100)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     687..708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             751..780
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 ggtaccgaaa acctgtactt ccaatccggt tctatggtga gcaagggcga ggagctgttc
       61 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      121 gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
      181 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
      241 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      301 cccgaaggct acgtccagga gcgcaccatc tccttcaagg acgacggcac ctacaagacc
      361 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      421 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac
      481 aacgtctata tcacggccga caagcagaag aacggcatca aggcgaactt caagatccgc
      541 cacaacgtcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      601 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
      661 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      721 atcactctcg gcatggacga gctgtacaag catcaccatc accatcacca tcaccatcac
      781 tag
//
