LOCUS       sequence-457679-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes an acceptor-only, TdLanYFP-tagged version of the LC3B 
            biosensor under the control of the CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  AUTHORS   Gokerkucuk EB, Cheron A, Tramier M, Bertolin G
  TITLE     The LC3B FRET biosensor monitors the modes of action of ATG4B during
            autophagy in living cells.
  JOURNAL   Autophagy. 2023 Aug;19(8):2275-2295. doi: 
            10.1080/15548627.2023.2179845. Epub 2023 Feb 22.
  PUBMED    36814061
REFERENCE   2  (bases 1 to 5752)
  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"; doi: 
            "10.1080/15548627.2023.2179845"; journalName: "Autophagy"; date: 
            "2023-08"; volume: "19"; issue: "8"; pages: "2275-2295"
COMMENT     Sequence Label: LC3B-TdLanYFP
FEATURES             Location/Qualifiers
     source          1..5752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..154
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             230..604
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
     CDS             638..1297
                     /codon_start=1
                     /product="tetrameric yellow fluorescent protein from 
                     Branchiostoma lanceolatum, also known as blFP-Y3"
                     /label=LanYFP
                     /translation="MASLPATHELHIFGSFNGVDFDMVGRGTGNPNDGYEELNLKSTKG
                     DLQFSPWILVPQIGYGFHQYLPFPDGMSPFQAAMKDGSGYQVHRTMQFEDGASLTSNYR
                     YTYEGSHIKGEFQVKGTGFPADGPVMTNSLTAADWCVTKMLYPNDKTIISTFDWTYTTG
                     NGKRYQSTARTTYTFAKPMAANILKNQPMFVFRKTELKHSKTELNFKEWQKAFTDVM"
     CDS             1349..2008
                     /codon_start=1
                     /product="tetrameric yellow fluorescent protein from 
                     Branchiostoma lanceolatum, also known as blFP-Y3"
                     /label=LanYFP
                     /translation="MASLPATHELHIFGSFNGVDFDMVGRGTGNPNDGYEELNLKSTKG
                     DLQFSPWILVPQIGYGFHQYLPFPDGMSPFQAAMKDGSGYQVHRTMQFEDGASLTSNYR
                     YTYEGSHIKGEFQVKGTGFPADGPVMTNSLTAADWCVTKMLYPNDKTIISTFDWTYTTG
                     NGKRYQSTARTTYTFAKPMAANILKNQPMFVFRKTELKHSKTELNFKEWQKAFTDVM"
     polyA_signal    2155..2276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2192..2211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2246..2265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2283..2738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2420..2441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2632..2651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2765..2869
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2871..3228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2871..3062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2894..2914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3079..3214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3263..4057
                     /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             3263..4057
                     /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(3317..3336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3927..3946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4245..4264)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4289..4336
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4665..5253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5154..5173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5428..5731
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5732..5752,1..183)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
       61 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      121 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      181 gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatcta tgccgtcgga
      241 gaagaccttc aagcagcgcc gcaccttcga acaaagagta gaagatgtcc gacttattcg
      301 agagcagcat ccaaccaaaa tcccggtgat aatagaacga tacaagggtg agaagcagct
      361 tcctgttctg gataaaacaa agttccttgt acctgaccat gtcaacatga gtgagctcat
      421 caagataatt agaaggcgct tacagctcaa tgctaatcag gccttcttcc tgttggtgaa
      481 cggacacagc atggtcagcg tctccacacc aatctcagag gtgtatgaga gtgagaaaga
      541 tgaagatgga ttcctgtaca tggtctacgc ctcccaggag acgttcggga tgaaattgtc
      601 agtggaattc atggtctcca aaggagagga ggataacatg gcctctctcc cagcgacaca
      661 tgagttacac atctttggct ccttcaacgg tgtggacttt gacatggtgg gtcgtggcac
      721 cggcaatcca aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca
      781 gttctccccc tggattctgg tccctcaaat cgggtatgga ttccatcagt acctgccctt
      841 ccccgacggg atgtcgcctt tccaggccgc catgaaagat ggctccggat accaagtcca
      901 tcgcacaatg cagtttgaag acggtgcctc cctgacttcc aactaccgct acacctacga
      961 gggaagccac atcaaaggag agtttcaggt gaaggggact ggtttccctg ctgacggtcc
     1021 tgtgatgacc aactcgctga ccgctgcgga ctggtgcgtg accaagatgc tgtaccccaa
     1081 cgacaaaacc atcatcagca cctttgactg gacttacacc actggaaatg gcaagcgcta
     1141 ccagagcact gcgcggacca cctacacctt tgccaagcca atggcggcca acatcctgaa
     1201 gaaccagccg atgttcgtgt tccgtaagac ggaactcaag cactccaaga ccgaactcaa
     1261 cttcaaggag tggcaaaagg catttaccga tgtgatgtcg actggcactg gttctacggg
     1321 ctcgggctcc tcaggagagg aggataacat ggcctctctc ccagcgacac atgagttaca
     1381 catctttggc tccttcaacg gtgtggactt tgacatggtg ggtcgtggca ccggcaatcc
     1441 aaatgatggt tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc
     1501 ctggattctg gtccctcaaa tcgggtatgg cttccatcag tacctgccct tccccgacgg
     1561 gatgtcgcct ttccaggccg ccatgaaaga tggctccgga taccaagtcc atcgcacaat
     1621 gcagtttgaa gacggtgcct ccctgacttc caactaccgc tacacctacg agggaagcca
     1681 catcaaagga gagtttcagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac
     1741 caactcgctg accgctgcgg actggtgcgt gaccaagatg ctgtacccca acgacaaaac
     1801 catcatcagc acctttgact ggacttacac cactggaaat ggcaagcgct accagagcac
     1861 tgcgcggacc acctacacct ttgccaagcc aatggcggcc aacatcctga agaaccagcc
     1921 gatgttcgtg ttccgtaaga cggaactcaa gcactccaag accgaactca acttcaagga
     1981 gtggcaaaag gcatttaccg atgtgatggg catggacgag ctgtacaagt aaagcggccg
     2041 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2101 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2281 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2341 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2401 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2461 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2521 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2581 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2641 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2701 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2761 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2821 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2881 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3241 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3301 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3361 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3421 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3481 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3541 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3601 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3661 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3721 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3781 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3841 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3901 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3961 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4021 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4081 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4141 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4201 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4261 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4321 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4381 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4441 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4501 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4801 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5281 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5341 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     5401 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     5461 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     5521 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5581 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5641 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5701 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt ac
//
