LOCUS       sequence-252986-        3738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone encoding human ATG3 with all 22 lysine residues 
            mutated to arginine.
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 allKR
FEATURES             Location/Qualifiers
     source          1..3738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(144..171)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(263..349)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     398..420
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     412..429
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     413..429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    445..544
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1493..1592)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1609..1628)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1610..1628)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1633..1649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1633..1649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2080..2871
                     /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      2964..3552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3453..3472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3706..3723
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
      121 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
      181 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
      241 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
      301 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
      361 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
      421 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
      481 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagca
      541 ggcttaatgc agaatgtgat taatactgtg aggggaaggg cactggaagt ggctgagtac
      601 ctgaccccgg tcctcaggga atcaaggttt agggaaacag gtgtaattac cccagaagag
      661 tttgtggcag ctggagatca cctagtccac cactgtccaa catggcaatg ggctacaggg
      721 gaagaattga gagtgagggc atacctacca acaggcagac aatttttggt aaccagaaat
      781 gtgccgtgct ataggcggtg cagacagatg gaatattcag atgaattgga agctatcatt
      841 gaagaagatg atggtgatgg cggatgggta gatacatatc acaacacagg tattacagga
      901 ataacggaag ccgttagaga gatcacactg gaaaataggg acaatataag gcttcaagat
      961 tgctcagcac tatgtgaaga ggaagaagat gaagatgaag gagaagctgc agatatggaa
     1021 gaatatgaag agagtggatt gttggaaaca gatgaggcta ccctagatac aaggagaata
     1081 gtagaagctt gtagagccag aactgatgct ggcggtgaag atgctatttt gcaaaccaga
     1141 acttatgacc tttacatcac ttatgataga tattaccaga ctccacgatt atggttgttt
     1201 ggctatgatg agcaacggca gcctttaaca gttgagcaca tgtatgaaga catcagtcag
     1261 gatcatgtga ggagaacagt gaccattgaa aatcaccctc atctgccacc acctcccatg
     1321 tgttcagttc acccatgcag gcatgctgag gtgatgagga gaatcattga gactgttgca
     1381 gaaggagggg gagaacttgg agttcatatg tatcttctta ttttcttgag atttgtacaa
     1441 gctgtcattc caacaataga atatgactac acaagacact tcacaatgta atacccagct
     1501 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1561 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1621 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1681 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
     1741 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1801 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     1861 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     1921 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     1981 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     2041 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     2101 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     2161 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     2221 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     2281 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     2341 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     2401 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     2461 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     2521 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2581 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2641 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2701 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2761 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2821 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     2881 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2941 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3001 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3061 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3121 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3181 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3241 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3301 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3361 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3421 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3481 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3541 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3601 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3661 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3721 gaggaagcgg aagagcgc
//
