LOCUS       sequence-244490-        3171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone encoding human MAP1LC3B2 wild-type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3171)
  AUTHORS   Agrotis A, Pengo N, Burden JJ, Ketteler R
  TITLE     Redundancy of human ATG4 protease isoforms in autophagy and 
            LC3/GABARAP processing revealed in cells.
  JOURNAL   Autophagy. 2019 Jan 21. doi: 10.1080/15548627.2019.1569925.
  PUBMED    30661429
REFERENCE   2  (bases 1 to 3171)
  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: "Autophagy. 
            2019 Jan 21. doi: 10.1080/15548627.2019.1569925."
COMMENT     Sequence Label: pDONR223 LC3B2 WT
FEATURES             Location/Qualifiers
     source          1..3171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(270..297)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(389..475)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     524..546
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     538..555
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     539..555
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    571..670
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             673..1047
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVCASQETFGMKLSV"
     protein_bind    complement(1052..1151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1168..1187)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1169..1187)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1192..1208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1192..1208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1639..2430
                     /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      2523..3111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3012..3031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
      241 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
      301 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
      361 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
      421 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
      481 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
      541 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg
      601 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
      661 aaagcaggct taatgccgtc ggagaagacc ttcaagcagc ggcgcacctt cgaacaaaga
      721 gtagaagatg tccgacttat tcgagagcag catccaacca aaatcccggt gataatagaa
      781 cgatacaagg gtgagaagca gcttcctgtt ctggataaaa caaagttcct tgtacctgac
      841 catgtcaaca tgagtgagct catcaagata attagaaggc gcttacagct caatgctaat
      901 caggccttct tcctgttggt gaacggacac agcatggtca gcgtctccac accaatctca
      961 gaggtgtatg agagtgagaa agatgaagat ggattcctgt acatggtctg tgcctcccag
     1021 gagacgttcg ggatgaaatt gtcagtgtaa tacccagctt tcttgtacaa agttggcatt
     1081 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1141 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     1201 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1261 aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta
     1321 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca
     1381 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
     1441 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     1501 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     1561 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     1621 aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca
     1681 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
     1741 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
     1801 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
     1861 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
     1921 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
     1981 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
     2041 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
     2101 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
     2161 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     2221 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     2281 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     2341 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     2401 ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt
     2461 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2521 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2581 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2641 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2701 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2761 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2821 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2881 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2941 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3001 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3061 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3121 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat g
//
