LOCUS       sequence-175204-        6325 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For  expression of the mammalian Atg8 protein LC3B without a tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6325)
  AUTHORS   Nguyen TN, Padman BS, Usher J, Oorschot V, Ramm G, Lazarou M
  TITLE     Atg8 family LC3/GABARAP proteins are crucial for 
            autophagosome-lysosome fusion but not autophagosome formation during
            PINK1/Parkin mitophagy and starvation.
  JOURNAL   J Cell Biol. 2016 Dec 19;215(6):857-874. Epub 2016 Nov 18.
  PUBMED    27864321
REFERENCE   2  (bases 1 to 6325)
  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 Cell 
            Biol."; date: "2016-12-19"; volume: "215(6)"; pages: "857-874. Epub 
            2016 Nov 18"
COMMENT     Sequence Label: pMX-IY LC3B
FEATURES             Location/Qualifiers
     source          1..6325
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    13..565
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(157..174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     384..403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(577..579,580..582,583..1296)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(622..643)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(883..902)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1230..1251
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1467..1936
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2066..2083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2237..2825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2317..2336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2989..3780,3781..3849))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3612..3631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3850..3954)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3984..4577
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4640..4997
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5056..5472
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5402..5424
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5440..5456
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5482..5856
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5794..5810
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             5910..6284
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
ORIGIN
        1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
       61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
      121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
      181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
      241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
      301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
      361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
      421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
      481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
      541 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg
      601 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      661 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      721 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
      781 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      841 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      901 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      961 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1021 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1081 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1141 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     1201 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1261 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acgataaaat aaaagatttt
     1321 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     1381 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     1441 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     1501 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     1561 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     1621 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     1681 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     1741 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     1801 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     1861 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     1921 cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta
     1981 tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2041 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2101 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2161 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2221 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2641 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc
     2941 cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3961 tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg caagctagct
     4021 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga aaagttcaga
     4081 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     4141 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     4201 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4261 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4321 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4381 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     4441 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4501 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4561 agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc cagggaccac
     4621 cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc gattgtctag
     4681 tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag ctctgtatct
     4741 ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct gggagacgtc
     4801 ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt
     4861 ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg agacgagaac
     4921 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc
     4981 gcgcgtcttg tctgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     5041 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     5101 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     5161 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     5221 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     5281 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     5341 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     5401 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     5461 tctctaggcg cccccatatg gccatatgag atcttatatg gggcaccccc gccccttgta
     5521 aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca agctcactta
     5581 caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc ctaccaagaa
     5641 caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt gtgggtccgc
     5701 cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt cctgctgacc
     5761 acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc ccacgtgaag
     5821 gctgccgacc ccgggggtgg accatcctct agactgccgg atctagctag ttaattaagg
     5881 atcccagtgt ggtggtacgg aattccacca tgccgtcgga gaagaccttc aagcagcgcc
     5941 gcaccttcga acaaagagta gaagatgtcc gacttattcg agagcagcat ccaaccaaaa
     6001 tcccggtgat aatagaacga tacaagggtg agaagcagct tcctgttctg gataaaacaa
     6061 agttccttgt acctgaccat gtcaacatga gtgagctcat caagataatt agaaggcgct
     6121 tacagctcaa tgctaatcag gccttcttcc tgttggtgaa cggacacagc atggtcagcg
     6181 tctccacacc aatctcagag gtgtatgaga gtgagaaaga tgaagatgga ttcctgtaca
     6241 tggtctacgc ctcccaggag acgttcggga tgaaattgtc agtgtaactc gagggccggc
     6301 gcgccgcggc cgctacgtaa attcc
//
