LOCUS Exported 6337 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Vimentin labeling with near-infrared fluorescent protein miRFP703. ACCESSION . VERSION . KEYWORDS pVimentin-miRFP703 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6337) AUTHORS Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, Verkhusha VV TITLE Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. JOURNAL Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405. PUBMED 27539380 REFERENCE 2 (bases 1 to 6337) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6337 /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 2050..2059 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2056..3003 /codon_start=1 /product="bright monomeric near-infrared fluorescent protein engineered from a bacterial phytochrome, with an emission maximum of 703 nm (Shcherbakova et al., 2016)" /label=miRFP703 /label=ORF frame 1 /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV VCHHYLPRFIRFELRAICKRLAERIATRITALES" polyA_signal 3125..3246 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3162..3181) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3216..3235 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3253..3708) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3390..3411) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3602..3621 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3735..3839 /gene="bla" /label=AmpR promoter promoter 3841..4198 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3864..3884) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4049..4184 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4111..4130 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4233..5027 /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(4287..4306) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4897..4916 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5215..5234) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5259..5306 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5635..6223 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6124..6143 /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 ccggactcag atctcgagct caagcttcga attcccatgt ccaccaggtc cgtgtcctcg 661 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg 721 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc 781 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg 841 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg 901 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag 961 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat 1021 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac 1081 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa 1141 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa 1201 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag 1261 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa 1321 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag 1381 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc 1441 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa 1501 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc 1561 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt 1621 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa 1681 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag 1741 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat 1801 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag 1861 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg 1921 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa 1981 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaaggggat 2041 ccaccggtcg ccaccatggt agcaggtcat gcctctggca gccccgcatt cgggaccgcc 2101 tctcattcga attgcgaaca tgaagagatc cacctcgccg gctcgatcca gccgcatggc 2161 gcgcttctgg tcgtcagcga acatgatcat cgcgtcatcc aggccagcgc caacgccgcg 2221 gaatttctga atctcggaag cgtactcggc gttccgctcg ccgagatcga cggcgatctg 2281 ttgatcaaga tcctgccgca tctcgatccc accgccgaag gcatgccggt cgcggtgcgc 2341 tgccggatcg gcaatccctc tacggagtac tgcggtctga tgcatcggcc tccggaaggc 2401 gggctgatca tcgaactcga acgtgccggc ccgtcgatcg atctgtcagg cacgctggcg 2461 ccggcgctgg agcggatccg cacggcgggt tcactgcgcg cgctgtgcga tgacaccgtg 2521 ctgctgtttc agcagtgcac cggctacgac cgggtgatgg tgtatcgttt cgatgagcaa 2581 ggccacggcc tggtattctc cgagtgccat gtgcctgggc tcgaatccta tttcggcaac 2641 cgctatccgt cgtcgctggt cccgcagatg gcgcggcagc tgtacgtgcg gcagcgcgtc 2701 cgcgtgctgg tcgacgtcac ctatcagccg gtgccgctgg agccgcggct gtcgccgctg 2761 accgggcgcg atctcgacat gtcgggctgc ttcctgcgct cgatgtcgcc gatccatctg 2821 cagttcctga aggacatggg cgtgcgcgcc accctggcgg tgtcgctggt ggtcggcggc 2881 aagctgtggg gcctggttgt ctgtcaccat tatctgccgc gcttcatccg tttcgagctg 2941 cgggcgatct gcaaacggct cgccgaaagg atcgcgacgc ggatcaccgc gcttgagagc 3001 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc 3061 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 3121 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3241 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 3301 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 3361 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 3421 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 3481 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 3541 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 3601 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 3661 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt 3721 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 3781 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc 3841 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 3901 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 3961 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 4021 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 4081 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 4141 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga 4201 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 4261 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 4321 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 4381 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc 4441 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4501 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4561 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4621 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4681 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4741 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg 4801 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4861 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4921 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4981 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 5041 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 5101 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 5161 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac 5221 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa 5281 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc 5341 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc 5401 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg 5461 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta 5521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 5581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 5641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 5701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 5761 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 5821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 5881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 5941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 6001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 6061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 6121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 6181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 6241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 6301 ccctgattct gtggataacc gtattaccgc catgcat //