LOCUS Exported 6335 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Vimentin fused to monomeric near-infrared fluorescent protein miRFP. ACCESSION . VERSION . KEYWORDS pmiRFP-Vimentin-17-C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6335) AUTHORS Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C, Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, Sabatini BL, Boyden ES TITLE A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters. JOURNAL Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 10.1038/s41589-018-0004-9. PUBMED 29483642 REFERENCE 2 (bases 1 to 6335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6335 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 606..615 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 3123..3244 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3160..3179) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3214..3233 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3251..3706) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3388..3409) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3600..3619 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3733..3837 /gene="bla" /label=AmpR promoter promoter 3839..4196 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3862..3882) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4047..4182 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4109..4128 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4231..5025 /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(4285..4304) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4895..4914 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5213..5232) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5257..5304 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5633..6221 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6122..6141 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtgccac catggtcgca ggacacgcaa gtggaagccc cgactttgga accgccgatc 661 cgagcgactg tgagagagaa gaaatccatc tggccggatc catccagcca cacggaacac 721 tgctggtggt ctccgagcct gaccatagaa tcattcaggc atctgccaac gccgctgaat 781 tcctgaatct gggaagtgtg ctgggagtcc ccctggctga gatcgacggg gatctgctga 841 tcaagattct gccccacctg gaccctaccg cagaaggaat gccagtggcc gtcaggtgcc 901 gcattggaaa cccctccaca gagtacgatg gactgatgca tcgaccacct gaaggaggac 961 tgatcattga gctggaacgg gctgggccac ccattgatct gagcggaaca ctggctcccg 1021 cactggagcg aattcgaact gcaggatccc tgagggctct gtgcgacgat actgcactgc 1081 tgttccagca gtgtaccggc tacgaccgag tgatggtcta tcggttcgat gaacaggggc 1141 acggagaggt gtattcagaa atacatgtga ctggactgga gagctacttt ggcaacaggt 1201 atcctagctc cctcgtgcca cagatggccc ggagactgta cgaaagacag agggtgcgcg 1261 tcctggtgga tgtctcttat cagcctgtgc cactggagcc aagactgagt cccctgactg 1321 gcagggacct ggatatgtca gggtgttttc tgcgctctat gagtcctact cacctgcagt 1381 tcctgaagaa tatgggcgtg cgggcaacac tggtggtcag cctggtggtc ggaggcaaac 1441 tgtgggggct ggtgatctgc caccattatc tgccacgctt catccatttt gagctgcgag 1501 caatttgtga gctgctggcc gaagctatcg caacccgcat tacagccctg agatctggca 1561 gcggtggagg cagcgcatcc ggcggaagcg gaagctccac caggtccgtg tcctcgtcct 1621 cctaccgcag gatgttcggc ggcccgggca ccgcgagccg gccgagctcc agccggagct 1681 acgtgactac gtccacccgc acctacagcc tgggcagcgc gctgcgcccc agcaccagcc 1741 gcagcctcta cgcctcgtcc ccgggcggcg tgtatgccac gcgctcctct gccgtgcgcc 1801 tgcggagcag cgtgcccggg gtgcggctcc tgcaggactc ggtggacttc tcgctggccg 1861 acgccatcaa caccgagttc aagaacaccc gcaccaacga gaaggtggag ctgcaggagc 1921 tgaatgaccg cttcgccaac tacatcgaca aggtgcgctt cctggagcag cagaataaga 1981 tcctgctggc cgagctcgag cagctcaagg gccaaggcaa gtcgcgcctg ggggacctct 2041 acgaggagga gatgcgggag ctgcgccggc aggtggacca gctaaccaac gacaaagccc 2101 gcgtcgaggt ggagcgcgac aacctggccg aggacatcat gcgcctccgg gagaaattgc 2161 aggaggagat gcttcagaga gaggaagccg aaaacaccct gcaatctttc agacaggatg 2221 ttgacaatgc gtctctggca cgtcttgacc ttgaacgcaa agtggaatct ttgcaagaag 2281 agattgcctt tttgaagaaa ctccacgaag aggaaatcca ggagctgcag gctcagattc 2341 aggaacagca tgtccaaatc gatgtggatg tttccaagcc tgacctcacg gctgccctgc 2401 gtgacgtacg tcagcaatat gaaagtgtgg ctgccaagaa cctgcaggag gcagaagaat 2461 ggtacaaatc caagtttgct gacctctctg aggctgccaa ccggaacaat gacgccctgc 2521 gccaggcaaa gcaggagtcc actgagtacc ggagacaggt gcagtccctc acctgtgaag 2581 tggatgccct taaaggaacc aatgagtccc tggaacgcca gatgcgtgaa atggaagaga 2641 actttgccgt tgaagctgct aactaccaag acactattgg ccgcctgcag gatgagattc 2701 agaatatgaa ggaggaaatg gctcgtcacc ttcgtgaata ccaagacctg ctcaatgtta 2761 agatggccct tgacattgag attgccacct acaggaagct gctggaaggc gaggagagca 2821 ggatttctct gcctcttcca aacttttcct ccctgaacct gagggaaact aatctggatt 2881 cactccctct ggttgatacc cactcaaaaa ggacacttct gattaagacg gttgaaacta 2941 gagatggaca ggttatcaac gaaacttctc agcatcacga tgaccttgaa taaggatcca 3001 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 3061 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 3121 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 3181 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 3241 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa 3301 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa 3361 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac 3421 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa 3481 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct 3541 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa 3601 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc 3661 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt 3721 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3781 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct 3841 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 4021 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 4081 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 4141 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc 4201 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 4261 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 4321 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 4381 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg 4441 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 4501 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 4561 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 4621 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 4681 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 4741 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 4801 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 4861 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 4921 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 4981 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 5041 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 5101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 5161 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac 5221 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 5281 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg 5341 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac 5401 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc 5461 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat 5521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 5581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 5641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 5701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 5761 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact 5821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 5941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 6001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 6061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 6121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 6181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 6241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 6301 ctgattctgt ggataaccgt attaccgcca tgcat //