LOCUS       sequence-211014-        4368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses BC2-tag-LC3B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4368)
  AUTHORS   Virant D, Traenkle B, Maier J, Kaiser PD, Bodenhofer M, Schmees C, 
            Vojnovic I, Pisak-Lukats B, Endesfelder U, Rothbauer U
  TITLE     A peptide tag-specific nanobody enables high-quality labeling for 
            dSTORM imaging.
  JOURNAL   Nat Commun. 2018 Mar 2;9(1):930. doi: 10.1038/s41467-018-03191-2.
  PUBMED    29500346
REFERENCE   2  (bases 1 to 4368)
  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: "Nat 
            Commun."; date: "2018-03-2"; volume: "9(1):930. doi"; pages: " 
            10.1038/s41467-018-03191-2"
COMMENT     Sequence Label: pBC2T-LC3B
FEATURES             Location/Qualifiers
     source          1..4368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        247..450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     401..421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             534..905
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="PSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
                     LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKDE
                     DGFLYMVYASQETFGMKLSV"
     polyA_signal    1038..1159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1075..1094)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1129..1148
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1166..1621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1303..1324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1515..1534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1648..1752
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1754..2111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1754..1945
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1777..1797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1962..2097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2146..2940
                     /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             2146..2940
                     /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(2200..2219)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2810..2829
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3128..3147)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3172..3219
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3548..4136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4037..4056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(4311..4368,1..246)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
       61 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      121 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      181 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      241 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      301 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      361 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      421 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      481 taccggtcgc caccatgcct gatcgcaagc gcgccgtgag ccattggcag cagccgtcgg
      541 agaagacctt caagcagcgc cgcaccttcg aacaaagagt agaagatgtc cgacttattc
      601 gagagcagca tccaaccaaa atcccggtga taatagaacg atacaagggt gagaagcagc
      661 ttcctgttct ggataaaaca aagttccttg tacctgacca tgtcaacatg agtgagctca
      721 tcaagataat tagaaggcgc ttacagctca atgctaatca ggccttcttc ctgttggtga
      781 acggacacag catggtcagc gtctccacac caatctcaga ggtgtatgag agtgagaaag
      841 atgaagatgg attcctgtac atggtctatg cctcccagga gacgttcggg atgaaattgt
      901 cagtgtaagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
      961 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1021 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1081 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1141 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1201 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1261 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1321 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1381 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1441 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1501 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1561 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1621 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1741 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     1801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     1861 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     1921 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     1981 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2041 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2101 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2161 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2221 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2281 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2341 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2401 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2461 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2521 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2581 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2641 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     2701 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2761 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2821 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2881 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2941 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3001 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3061 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3121 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3181 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3241 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3301 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3361 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     3421 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3481 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3541 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3601 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3661 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3721 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3781 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3841 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3901 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3961 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4021 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4081 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4141 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4201 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     4261 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4321 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgccca
//
