LOCUS       Exported                5396 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses vimentin-BC2-tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pVIM-BC2T
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5396)
  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 5396)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5396
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             2059..2094
                     /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    2242..2363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2279..2298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2333..2352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2370..2825)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2507..2528)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2719..2738
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2852..2956
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2958..3315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2981..3001)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3166..3301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3228..3247
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3350..4144
                     /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(3404..3423)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4014..4033
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4332..4351)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4376..4423
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4752..5340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5241..5260
                     /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 tagcgccacc
      661 atgtccacca ggtccgtgtc ctcgtcctcc taccgcagga tgttcggcgg cccgggcacc
      721 gcgagccggc cgagctccag ccggagctac gtgactacgt ccacccgcac ctacagcctg
      781 ggcagcgcgc tgcgccccag caccagccgc agcctctacg cctcgtcccc gggcggcgtg
      841 tatgccacgc gctcctctgc cgtgcgcctg cggagcagcg tgcccggggt gcggctcctg
      901 caggactcgg tggacttctc gctggccgac gccatcaaca ccgagttcaa gaacacccgc
      961 accaacgaga aggtggagct gcaggagctg aatgaccgct tcgccaacta catcgacaag
     1021 gtgcgcttcc tggagcagca gaataagatc ctgctggccg agctcgagca gctcaagggc
     1081 caaggcaagt cgcgcctggg ggacctctac gaggaggaga tgcgggagct gcgccggcag
     1141 gtggaccagc taaccaacga caaagcccgc gtcgaggtgg agcgcgacaa cctggccgag
     1201 gacatcatgc gcctccggga gaaattgcag gaggagatgc ttcagagaga ggaagccgaa
     1261 aacaccctgc aatctttcag acaggatgtt gacaatgcgt ctctggcacg tcttgacctt
     1321 gaacgcaaag tggaatcttt gcaagaagag attgcctttt tgaagaaact ccacgaagag
     1381 gaaatccagg agctgcaggc tcagattcag gaacagcatg tccaaatcga tgtggatgtt
     1441 tccaagcctg acctcacggc tgccctgcgt gacgtacgtc agcaatatga aagtgtggct
     1501 gccaagaacc tgcaggaggc agaagaatgg tacaaatcca agtttgctga cctctctgag
     1561 gctgccaacc ggaacaatga cgccctgcgc caggcaaagc aggagtccac tgagtaccgg
     1621 agacaggtgc agtccctcac ctgtgaagtg gatgccctta aaggaaccaa tgagtccctg
     1681 gaacgccaga tgcgtgaaat ggaagagaac tttgccgttg aagctgctaa ctaccaagac
     1741 actattggcc gcctgcagga tgagattcag aatatgaagg aggaaatggc tcgtcacctt
     1801 cgtgaatacc aagacctgct caatgttaag atggcccttg acattgagat tgccacctac
     1861 aggaagctgc tggaaggcga ggagagcagg atttctctgc ctcttccaaa cttttcctcc
     1921 ctgaacctga gggaaactaa tctggattca ctccctctgg ttgataccca ctcaaaaagg
     1981 acacttctga ttaagacggt tgaaactaga gatggacagg ttatcaacga aacttctcag
     2041 catcacgatg accttgaacc agacagaaaa gcggctgtta gtcactggca gcaatagggt
     2101 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     2161 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2221 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2341 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     2401 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2461 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2521 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2581 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2641 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2701 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2761 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2821 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2881 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2941 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3241 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3301 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3361 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3421 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3481 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3541 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3601 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3661 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3721 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3781 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3841 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3901 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3961 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4021 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4081 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4141 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4201 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4261 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4321 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4381 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4441 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4501 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4561 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctca
//
