LOCUS       sequence-175205-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For  expression of the mammalian Atg8 protein LC3B without a tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  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 6283)
  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..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(44..835,836..904))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     667..686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(905..1009)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1024..1617
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1680..2037
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2102..2518
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2448..2470
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2486..2502
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2528..2902
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2840..2856
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2956..3330
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
     misc_feature    join(3373..3946,3947..3949,3950..3958)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3540..3557)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3767..3786
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3959..3961,3962..3964,3965..4678)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4004..4025)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4265..4284)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4612..4633
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4745..5338
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5404..5421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5575..6163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5655..5674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
       61 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
      661 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
      721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
      781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
      841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcctg
     1021 cataatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     1081 ggcatggaaa aatacataac tgagaataga aaagttcaga tcaaggtcag gaacagatgg
     1141 aacagctgaa tatgggccaa agcggatatc tgtggtaagc agttcctgcc ccggctcagg
     1201 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     1261 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     1321 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     1381 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     1441 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     1501 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     1561 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     1621 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     1681 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
     1741 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
     1801 gacgagttcg gaacacccgg ccgcaaccct gggagacgtc ccagggatct cgggggccgt
     1861 ttttgtggcc cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc
     1921 cttagaggag ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc
     1981 gtctgaattt ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag
     2041 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa atatgggccc
     2101 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     2161 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     2221 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     2281 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     2341 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     2401 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     2461 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc
     2521 catatggcca tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc
     2581 tgacatgaca agagttacta acagcccctc tctccaagct cacttacagg ctctctactt
     2641 agtccagcac gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc
     2701 ggtggtacct cacccttacc gagtcggcga cacagtgtgg gtccgccgac accagactaa
     2761 gaacctagaa cctcgctgga aaggacctta cacagtcctg ctgaccaccc ccaccgccct
     2821 caaagtagac ggcatcgcag cttggataca cgccgcccac gtgaaggctg ccgaccccgg
     2881 gggtggacca tcctctagac tgccggatct agctagttaa ttaaggatcc cagtgtggtg
     2941 gtacggaatt ccaccatgcc gtcggagaag accttcaagc agcgccgcac cttcgaacaa
     3001 agagtagaag atgtccgact tattcgagag cagcatccaa ccaaaatccc ggtgataata
     3061 gaacgataca agggtgagaa gcagcttcct gttctggata aaacaaagtt ccttgtacct
     3121 gaccatgtca acatgagtga gctcatcaag ataattagaa ggcgcttaca gctcaatgct
     3181 aatcaggcct tcttcctgtt ggtgaacgga cacagcatgg tcagcgtctc cacaccaatc
     3241 tcagaggtgt atgagagtga gaaagatgaa gatggattcc tgtacatggt ctatgcctcc
     3301 caggagacgt tcgggatgaa attgtcagtg taactcgagg gccggcgcgc cgcggccgct
     3361 acgtaaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     3421 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3481 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3541 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3601 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3661 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3721 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3781 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3841 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc
     3901 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccat
     3961 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4141 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     4201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4381 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4441 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4561 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     4681 cggcccagcc acagtggtcg acgataaaat aaaagatttt atttagtctc cagaaaaagg
     4741 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgga
     4801 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     4861 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4921 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4981 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     5041 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     5101 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     5161 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     5221 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     5281 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcattct
     5341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6001 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttgcggccg gcc
//
