LOCUS       sequence-215722-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For  expression of the mammalian Atg8 protein LC3C without a tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  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 6412)
  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 LC3C
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             82..675
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    738..1095
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1154..1570
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1500..1522
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1538..1554
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1580..1954
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1892..1908
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    join(2500..3074,3075..3077,3078..3086)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2667..2684)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2894..2913
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3087..3089,3090..3092,3093..3806)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3132..3153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3393..3412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3740..3761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3977..4446
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4576..4593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4747..5335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4827..4846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5499..6290,6291..6359))
                     /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     6122..6141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6360..6412,1..52))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
       61 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
      121 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      181 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      241 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      301 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      361 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      421 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      481 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      541 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      601 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      661 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      721 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      781 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      841 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      901 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      961 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
     1021 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1081 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1141 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1201 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1261 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1321 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1381 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1441 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1501 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1561 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1621 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1681 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1741 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1801 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1861 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1921 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1981 cccagtgtgg tggtacggaa ttccaccatg ccgcctccac agaaaatccc aagcgtcaga
     2041 cccttcaagc agaggaaaag cttggcaatc agacaagagg aagttgctgg aatccgggca
     2101 aagttcccca acaaaatccc ggtggtagtg gagcgctacc ccagggagac gttcctgccc
     2161 ccgctggaca aaaccaagtt cctggtcccg caggagctga ccatgaccca gttcctcagc
     2221 atcatccgga gccgcatggt cctgagagcc acggaagcct tttacttgct ggtgaacaac
     2281 aagagcctgg tcagcatgag cgcaaccatg gcagagatct acagagacta caaggatgag
     2341 gatggcttcg tgtacatgac ctacgcctcc caggagacat ttggctgcct ggagtcagca
     2401 gcccccaggg atgggagcag ccttgaggac agaccctgca atcctctcta gctcgagggc
     2461 cggcgcgccg cggccgctac gtaaattccg cccccccccc ccctctccct cccccccccc
     2521 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2581 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2641 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2701 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2761 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2821 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2881 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2941 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     3001 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3061 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3121 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3181 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3241 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct
     3301 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3361 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3421 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3481 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3661 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     3721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3781 tcggcatgga cgagctgtac aagtaagtcg acgataaaat aaaagatttt atttagtctc
     3841 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     3901 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca
     3961 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4021 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     4081 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     4141 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     4201 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4261 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     4321 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     4381 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     4441 ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa
     4501 atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa
     5461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat ga
//
