LOCUS       sequence-490778-        5449 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Autophagosome marker LC3 tagged N-terminally with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5449)
  AUTHORS   Basak B, Holzbaur ELF
  TITLE     Mitochondrial damage triggers the concerted degradation of negative 
            regulators of neuronal autophagy.
  JOURNAL   Nat Commun. 2025 Aug 9;16(1):7367. doi: 10.1038/s41467-025-62379-5.
  PUBMED    40783388
REFERENCE   2  (bases 1 to 5449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-08-9"; volume: "16(1):7367. doi"; pages: " 
            10.1038/s41467-025-62379-5"
COMMENT     Sequence Label: mCherry-LC3
FEATURES             Location/Qualifiers
     source          1..5449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     545..564
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(653..1010)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      667..802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        819..1010
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     967..987
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1012..1116)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1143..1598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1230..1249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1440..1461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1605..1726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1616..1635)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1670..1689
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2196..2618)
                     /codon_start=1
                     /gene="Map1lc3b"
                     /product="rat LC3 protein"
                     /label=LC3
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
                     DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
     CDS             complement(2637..3344)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2912..2931)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3181..3199
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3341..3350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3395..3598)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3424..3444)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3599..3902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4077..4665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4157..4176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4994..5041
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5066..5085
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(5273..5449,1..618))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(join(5273..5449,1..618))
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5384..5403)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
       61 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
      121 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
      181 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
      241 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      301 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      361 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      421 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      481 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      541 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      601 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
      661 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
      721 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      781 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      841 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      901 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      961 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
     1021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1141 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1561 taacgcgaat tttaacaaaa tattaacgct tacaatttac gcgttaagat acattgatga
     1621 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1681 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1741 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1801 cctctacaaa tgtggtatgg ctgattatga tcagttatct agatccggtg gatcccgggc
     1861 ccgcggtacc gtcgactgca gaattcgcac catagttata aaccagggac atgacgacgt
     1921 acacaaccca cacacggcag cagtggggac ttacacagtg taacataaca ttgggggtta
     1981 gcagggacag ctgcatgcag ggagggtggg tgctgtgggg agccaggtcg agaggtggct
     2041 gtgttggctt ccgtccgtga cagaggcagc tctcactgag attctcagac acgagtgagt
     2101 gagtgaccgg ggatggtctg agtgtcacag tgggctccat gcaggtagca gggagcagag
     2161 gcttgcttta gttggaagtg gctgtatgtc tgtcacaagc atggctctct tcctgttgct
     2221 gttgccttca gagctgacat gtatgtaaca gccagtgctg tcccgaacgt ctcctgggag
     2281 gcatagacca tgtacaggaa gccgtcttca tctctctcgc tctcgtacac ttcagagatg
     2341 ggtgtggaca cactcaccat gctgtgccca ttcaccagga ggaagaaggc ttggttagca
     2401 ttgagctgca ggcgccttct aattatcttg atgagttcgc tcatattcac gtgatcaggt
     2461 acaaggaact tggtcttgtc caggacgggc agctgcttct cacccttgta tcgctctata
     2521 atcactggga tcttggtggg gtgctgctcc cggatgagcc ggacatcttc cactctttgt
     2581 tcgaagctcc ggcgctgttt gaaggtcttc tcggacggtc tagatctgag tccggacttg
     2641 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg
     2701 atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg
     2761 ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg
     2821 ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac
     2881 atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag
     2941 gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag
     3001 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag
     3061 ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc
     3121 ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc
     3181 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
     3241 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     3301 tccttgatga tggccatgtt atcctcctcg cccttgctca ccatggtggc aagcttaagt
     3361 ttaaacgcta gcggatctga cggttcacta aaccagctct gcttatatag acctcccacc
     3421 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     3481 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     3541 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     3601 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     3661 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     3721 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     3781 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     3841 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     3901 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     3961 taatgcatgg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4801 atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct
     4861 gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg
     4921 gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga
     4981 accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc
     5041 gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta
     5101 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     5161 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     5221 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     5281 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     5341 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     5401 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgat
//
