LOCUS       sequence-213817-        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        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..1378
                     /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(816..834)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1084..1103
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2810..2845
                     /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    2993..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3030..3049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3084..3103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3121..3576)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3258..3279)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3470..3489
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3603..3707
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3709..4066
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3709..3900
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3732..3752)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3917..4052
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4101..4895
                     /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             4101..4895
                     /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(4155..4174)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4765..4784
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5083..5102)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5127..5174
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5503..6091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5992..6011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtcagca agggcgagga ggataacatg gccatcatca aggagttcat
      721 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
      781 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
      841 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
      901 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
      961 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1021 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1081 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     1141 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     1201 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1261 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1321 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagca
     1381 gatctcgagc tcaagcttag gtggaggagg ttcttccacc aggtccgtgt cctcgtcctc
     1441 ctaccgcagg atgttcggcg gcccgggcac cgcgagccgg ccgagctcca gccggagcta
     1501 cgtgactacg tccacccgca cctacagcct gggcagcgcg ctgcgcccca gcaccagccg
     1561 cagcctctac gcctcgtccc cgggcggcgt gtatgccacg cgctcctctg ccgtgcgcct
     1621 gcggagcagc gtgcccgggg tgcggctcct gcaggactcg gtggacttct cgctggccga
     1681 cgccatcaac accgagttca agaacacccg caccaacgag aaggtggagc tgcaggagct
     1741 gaatgaccgc ttcgccaact acatcgacaa ggtgcgcttc ctggagcagc agaataagat
     1801 cctgctggcc gagctcgagc agctcaaggg ccaaggcaag tcgcgcctgg gggacctcta
     1861 cgaggaggag atgcgggagc tgcgccggca ggtggaccag ctaaccaacg acaaagcccg
     1921 cgtcgaggtg gagcgcgaca acctggccga ggacatcatg cgcctccggg agaaattgca
     1981 ggaggagatg cttcagagag aggaagccga aaacaccctg caatctttca gacaggatgt
     2041 tgacaatgcg tctctggcac gtcttgacct tgaacgcaaa gtggaatctt tgcaagaaga
     2101 gattgccttt ttgaagaaac tccacgaaga ggaaatccag gagctgcagg ctcagattca
     2161 ggaacagcat gtccaaatcg atgtggatgt ttccaagcct gacctcacgg ctgccctgcg
     2221 tgacgtacgt cagcaatatg aaagtgtggc tgccaagaac ctgcaggagg cagaagaatg
     2281 gtacaaatcc aagtttgctg acctctctga ggctgccaac cggaacaatg acgccctgcg
     2341 ccaggcaaag caggagtcca ctgagtaccg gagacaggtg cagtccctca cctgtgaagt
     2401 ggatgccctt aaaggaacca atgagtccct ggaacgccag atgcgtgaaa tggaagagaa
     2461 ctttgccgtt gaagctgcta actaccaaga cactattggc cgcctgcagg atgagattca
     2521 gaatatgaag gaggaaatgg ctcgtcacct tcgtgaatac caagacctgc tcaatgttaa
     2581 gatggccctt gacattgaga ttgccaccta caggaagctg ctggaaggcg aggagagcag
     2641 gatttctctg cctcttccaa acttttcctc cctgaacctg agggaaacta atctggattc
     2701 actccctctg gttgataccc actcaaaaag gacacttctg attaagacgg ttgaaactag
     2761 agatggacag gttatcaacg aaacttctca gcatcacgat gaccttgaac cagacagaaa
     2821 agcggctgtt agtcactggc agcaataggg taccgcgggc ccgggatcca ccggatctag
     2881 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2941 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3121 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3181 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3241 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3301 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3361 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3421 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3481 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3541 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3601 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3661 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3721 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3901 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3961 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4021 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4081 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4141 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4201 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4261 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4321 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4381 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4441 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4501 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4561 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4621 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4681 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4741 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4801 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4861 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4921 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4981 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5041 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5101 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5161 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5221 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5281 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5341 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5401 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5461 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5521 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5581 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5641 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5701 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5761 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5821 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5881 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5941 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6001 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6061 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6121 cctggccttt tgctggcctt ttgctca
//
