LOCUS       sequence-244529-        3156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone encoding human MAP1LC3A lacking stop codon, 
            suitable for C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3156)
  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 3156)
  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 LC3A no-stop
FEATURES             Location/Qualifiers
     source          1..3156
                     /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..1035
                     /codon_start=1
                     /gene="MAP1LC3A"
                     /product="human LC3 protein"
                     /label=LC3A
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSDRPFKQRRSFADRCKEVQQIRDQHPSKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELVKIIRRRLQLNPTQAFFLLVNQHSMVSVSTPIADIYEQEKD
                     EDGFLYMVYASQETFGF"
     RBS             723..731
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(1037..1136)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1153..1172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1154..1172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1177..1193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1177..1193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1624..2415
                     /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      2508..3096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2997..3016
                     /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 taatgccctc agaccggcct ttcaagcagc ggcggagctt cgccgaccgc
      721 tgtaaggagg tacagcagat ccgcgaccag caccccagca aaatcccggt gatcatcgag
      781 cgctacaagg gtgagaagca gctgcccgtc ctggacaaga ccaagttttt ggtcccggac
      841 catgtcaaca tgagcgagtt ggtcaagatc atccggcgcc gcctgcagct gaaccccacg
      901 caggccttct tcctgctggt gaaccagcac agcatggtga gtgtgtccac gcccatcgcg
      961 gacatctacg agcaggagaa agacgaggac ggcttcctct atatggtcta cgcctcccag
     1021 gaaaccttcg gcttctaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1081 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1141 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1201 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatgc
     1261 ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg
     1321 tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt
     1381 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
     1441 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
     1501 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
     1561 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
     1621 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1681 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1741 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1801 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     1861 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     1921 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     1981 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     2041 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     2101 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     2161 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     2221 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     2281 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     2341 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     2401 gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg
     2461 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2521 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2581 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2641 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2701 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2761 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2821 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2881 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2941 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3001 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3061 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3121 cggttcctgg ccttttgctg gccttttgct cacatg
//
