LOCUS       sequence-252979-        3738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone encoding human ATG3 C264A (catalytic inactive).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3738)
  AUTHORS   Agrotis A, von Chamier L, Oliver H, Kiso K, Singh T, Ketteler R
  TITLE     Human ATG4 autophagy proteases counteract attachment of 
            ubiquitin-like LC3/GABARAP proteins to other cellular proteins.
  JOURNAL   J Biol Chem. 2019 Jul 17. pii: AC119.009977. doi: 
            10.1074/jbc.AC119.009977.
  PUBMED    31315929
REFERENCE   2  (bases 1 to 3738)
  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: "J Biol 
            Chem. 2019 Jul 17. pii: AC119.009977. doi: 
            10.1074/jbc.AC119.009977."
COMMENT     Sequence Label: pDONR223 ATG3 C264A
FEATURES             Location/Qualifiers
     source          1..3738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..57
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(216..243)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(335..421)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     470..492
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     484..501
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     485..501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    517..616
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1565..1664)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1681..1700)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1682..1700)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1705..1721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1705..1721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2152..2943
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3036..3624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3525..3544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
       61 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
      181 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
      241 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
      301 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
      361 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
      421 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
      481 cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg
      541 atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg
      601 tacaaaaaag caggcttaat gcagaatgtg attaatactg tgaagggaaa ggcactggaa
      661 gtggctgagt acctgacccc ggtcctcaag gaatcaaagt ttaaggaaac aggtgtaatt
      721 accccagaag agtttgtggc agctggagat cacctagtcc accactgtcc aacatggcaa
      781 tgggctacag gggaagaatt gaaagtgaag gcatacctac caacaggcaa acaatttttg
      841 gtaaccaaaa atgtgccgtg ctataagcgg tgcaaacaga tggaatattc agatgaattg
      901 gaagctatca ttgaagaaga tgatggtgat ggcggatggg tagatacata tcacaacaca
      961 ggtattacag gaataacgga agccgttaaa gagatcacac tggaaaataa ggacaatata
     1021 aggcttcaag attgctcagc actatgtgaa gaggaagaag atgaagatga aggagaagct
     1081 gcagatatgg aagaatatga agagagtgga ttgttggaaa cagatgaggc taccctagat
     1141 acaaggaaaa tagtagaagc ttgtaaagcc aaaactgatg ctggcggtga agatgctatt
     1201 ttgcaaacca gaacttatga cctttacatc acttatgata aatattacca gactccacga
     1261 ttatggttgt ttggctatga tgagcaacgg cagcctttaa cagttgagca catgtatgaa
     1321 gacatcagtc aggatcatgt gaagaaaaca gtgaccattg aaaatcaccc tcatctgcca
     1381 ccacctccca tgtgttcagt tcacccagcc aggcatgctg aggtgatgaa gaaaatcatt
     1441 gagactgttg cagaaggagg gggagaactt ggagttcata tgtatcttct tattttcttg
     1501 aaatttgtac aagctgtcat tccaacaata gaatatgact acacaagaca cttcacaatg
     1561 taatacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     1621 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     1681 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1741 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc
     1801 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
     1861 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     1921 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     1981 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     2041 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     2101 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     2161 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     2221 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     2281 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     2341 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     2401 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     2461 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     2521 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2581 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2641 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2701 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2761 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2821 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2881 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2941 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3181 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3421 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3661 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3721 cgtattaccg cctttgag
//
