LOCUS       sequence-210840-        6147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mCherry-vimentin-BC2-tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6147)
  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 6147)
  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: pmCherry-VIM-BC2T
FEATURES             Location/Qualifiers
     source          1..6147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             605..1312
                     /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(750..768)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1018..1037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2744..2779
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     polyA_signal    2927..3048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2964..2983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3018..3037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3055..3510)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3192..3213)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3404..3423
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3537..3641
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3643..4000
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3643..3834
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3666..3686)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3851..3986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4035..4829
                     /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             4035..4829
                     /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(4089..4108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4699..4718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5017..5036)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5061..5108
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5437..6025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5926..5945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      601 caccatggtc agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
      661 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      721 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      781 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
      841 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
      901 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
      961 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1021 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     1081 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     1141 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1201 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1261 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agcagatctc
     1321 gagctcaagc ttaggtggag gaggttcttc caccaggtcc gtgtcctcgt cctcctaccg
     1381 caggatgttc ggcggcccgg gcaccgcgag ccggccgagc tccagccgga gctacgtgac
     1441 tacgtccacc cgcacctaca gcctgggcag cgcgctgcgc cccagcacca gccgcagcct
     1501 ctacgcctcg tccccgggcg gcgtgtatgc cacgcgctcc tctgccgtgc gcctgcggag
     1561 cagcgtgccc ggggtgcggc tcctgcagga ctcggtggac ttctcgctgg ccgacgccat
     1621 caacaccgag ttcaagaaca cccgcaccaa cgagaaggtg gagctgcagg agctgaatga
     1681 ccgcttcgcc aactacatcg acaaggtgcg cttcctggag cagcagaata agatcctgct
     1741 ggccgagctc gagcagctca agggccaagg caagtcgcgc ctgggggacc tctacgagga
     1801 ggagatgcgg gagctgcgcc ggcaggtgga ccagctaacc aacgacaaag cccgcgtcga
     1861 ggtggagcgc gacaacctgg ccgaggacat catgcgcctc cgggagaaat tgcaggagga
     1921 gatgcttcag agagaggaag ccgaaaacac cctgcaatct ttcagacagg atgttgacaa
     1981 tgcgtctctg gcacgtcttg accttgaacg caaagtggaa tctttgcaag aagagattgc
     2041 ctttttgaag aaactccacg aagaggaaat ccaggagctg caggctcaga ttcaggaaca
     2101 gcatgtccaa atcgatgtgg atgtttccaa gcctgacctc acggctgccc tgcgtgacgt
     2161 acgtcagcaa tatgaaagtg tggctgccaa gaacctgcag gaggcagaag aatggtacaa
     2221 atccaagttt gctgacctct ctgaggctgc caaccggaac aatgacgccc tgcgccaggc
     2281 aaagcaggag tccactgagt accggagaca ggtgcagtcc ctcacctgtg aagtggatgc
     2341 ccttaaagga accaatgagt ccctggaacg ccagatgcgt gaaatggaag agaactttgc
     2401 cgttgaagct gctaactacc aagacactat tggccgcctg caggatgaga ttcagaatat
     2461 gaaggaggaa atggctcgtc accttcgtga ataccaagac ctgctcaatg ttaagatggc
     2521 ccttgacatt gagattgcca cctacaggaa gctgctggaa ggcgaggaga gcaggatttc
     2581 tctgcctctt ccaaactttt cctccctgaa cctgagggaa actaatctgg attcactccc
     2641 tctggttgat acccactcaa aaaggacact tctgattaag acggttgaaa ctagagatgg
     2701 acaggttatc aacgaaactt ctcagcatca cgatgacctt gaaccagaca gaaaagcggc
     2761 tgttagtcac tggcagcaat agggtaccgc gggcccggga tccaccggat ctagataact
     2821 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2881 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2941 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3061 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3121 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3181 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3241 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3301 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3361 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3421 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3481 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3661 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3841 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3901 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3961 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4021 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4081 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4141 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4201 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4261 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4321 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4381 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4441 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4501 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4561 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4621 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4681 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4741 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4801 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4861 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4981 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5041 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5101 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5161 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5221 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5581 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6121 ccgtattacc gccatgcatt agttatt
//
