LOCUS       Exported                3156 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Gateway entry clone encoding human deconjugation-resistant LC3B.
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223 LC3B Q116P G120
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3156)
  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 3156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3156
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     332..348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     332..348
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     353..372
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        353..371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    389..488
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(855..954)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(970..987)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(970..986)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(979..1001)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1050..1136
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1228..1255
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1414..1431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1585..2173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1665..1684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2266..3057)
                     /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"
ORIGIN
        1 ggatgcccga ggcatagact gtaccccaaa aaaacagtca taacaagcca tgaaaaccgc
       61 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
      121 acgctacttg cattacagct tacgaaccga acaggcttat gtccactggg ttcgtgcctt
      181 catccgtttc cacggtgtgc gtcacccggc aaccttgggt agcagcgaag tcgaggcatt
      241 tctgtcctgg ctggtctaga ggaggcatga tgatatattt ttatcttgtg caatgtaaca
      301 tcagagattt tgagacacgg gccagagctg ccaggaaaca gctatgacca tgtaatacga
      361 ctcactatag gggatatcag ctggatggca aataatgatt ttattttgac tgatagtgac
      421 ctgttcgttg caacaaattg ataagcaatg ctttcttata atgccaactt tgtacaagaa
      481 agctgggtat tacccgaacg tctccgggga ggcatagacc atgtacagga atccatcttc
      541 atctttctca ctctcataca cctctgagat tggtgtggag acgctgacca tgctgtgtcc
      601 gttcaccaac aggaagaagg cctgattagc attgagctgt aagcgccttc taattatctt
      661 gatgagctca ctcatgttga catggtcagg tacaaggaac tttgttttat ccagaacagg
      721 aagctgcttc tcacccttgt atcgttctat tatcaccggg attttggttg gatgctgctc
      781 tcgaataagt cggacatctt ctactctttg ttcgaaggtg cgccgctgct tgaaggtctt
      841 ctccgacggc attaagcctg cttttttgta caaagttggc attataaaaa agcattgctc
      901 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
      961 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
     1021 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     1081 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1141 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1201 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     1261 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc
     1321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1381 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1441 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1501 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2221 cgcgtaactc acgttaaggg attttggtca tgggtggctc gagggttatt tgccgactac
     2281 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     2341 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     2401 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     2461 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     2521 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     2581 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     2641 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     2701 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     2761 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     2821 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     2881 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     2941 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     3001 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     3061 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     3121 caaacatcga cccacggcgt aacgcgcttg ctgctt
//
