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 49..352 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 353..556 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 507..527 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1989..2024 /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 2172..2293 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2209..2228) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2263..2282 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2300..2755) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2437..2458) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2649..2668 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2782..2886 /gene="bla" /label=AmpR promoter promoter 2888..3245 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2911..2931) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3096..3231 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3158..3177 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3280..4074 /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(3334..3353) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3944..3963 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4262..4281) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4306..4353 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4682..5270 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5171..5190 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgccacc atgtccacca 601 ggtccgtgtc ctcgtcctcc taccgcagga tgttcggcgg cccgggcacc gcgagccggc 661 cgagctccag ccggagctac gtgactacgt ccacccgcac ctacagcctg ggcagcgcgc 721 tgcgccccag caccagccgc agcctctacg cctcgtcccc gggcggcgtg tatgccacgc 781 gctcctctgc cgtgcgcctg cggagcagcg tgcccggggt gcggctcctg caggactcgg 841 tggacttctc gctggccgac gccatcaaca ccgagttcaa gaacacccgc accaacgaga 901 aggtggagct gcaggagctg aatgaccgct tcgccaacta catcgacaag gtgcgcttcc 961 tggagcagca gaataagatc ctgctggccg agctcgagca gctcaagggc caaggcaagt 1021 cgcgcctggg ggacctctac gaggaggaga tgcgggagct gcgccggcag gtggaccagc 1081 taaccaacga caaagcccgc gtcgaggtgg agcgcgacaa cctggccgag gacatcatgc 1141 gcctccggga gaaattgcag gaggagatgc ttcagagaga ggaagccgaa aacaccctgc 1201 aatctttcag acaggatgtt gacaatgcgt ctctggcacg tcttgacctt gaacgcaaag 1261 tggaatcttt gcaagaagag attgcctttt tgaagaaact ccacgaagag gaaatccagg 1321 agctgcaggc tcagattcag gaacagcatg tccaaatcga tgtggatgtt tccaagcctg 1381 acctcacggc tgccctgcgt gacgtacgtc agcaatatga aagtgtggct gccaagaacc 1441 tgcaggaggc agaagaatgg tacaaatcca agtttgctga cctctctgag gctgccaacc 1501 ggaacaatga cgccctgcgc caggcaaagc aggagtccac tgagtaccgg agacaggtgc 1561 agtccctcac ctgtgaagtg gatgccctta aaggaaccaa tgagtccctg gaacgccaga 1621 tgcgtgaaat ggaagagaac tttgccgttg aagctgctaa ctaccaagac actattggcc 1681 gcctgcagga tgagattcag aatatgaagg aggaaatggc tcgtcacctt cgtgaatacc 1741 aagacctgct caatgttaag atggcccttg acattgagat tgccacctac aggaagctgc 1801 tggaaggcga ggagagcagg atttctctgc ctcttccaaa cttttcctcc ctgaacctga 1861 gggaaactaa tctggattca ctccctctgg ttgataccca ctcaaaaagg acacttctga 1921 ttaagacggt tgaaactaga gatggacagg ttatcaacga aacttctcag catcacgatg 1981 accttgaacc agacagaaaa gcggctgtta gtcactggca gcaatagggt accgcgggcc 2041 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt 2101 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 2161 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 2221 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 2281 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 2341 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 2401 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 2461 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 2521 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 2581 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 2641 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 2701 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt 2761 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 2821 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 2881 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 2941 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3061 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3181 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3241 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 3301 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 3361 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3421 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta 3481 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3541 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3601 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3661 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3721 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3781 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc 3841 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 3901 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 3961 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4021 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 4081 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 4141 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 4201 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa 4261 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 4321 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 4381 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 4441 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 4501 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc 4561 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 4621 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 4681 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 4741 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 4801 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 4861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 4921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 4981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 5041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 5101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 5161 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 5221 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 5281 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 5341 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaata //