LOCUS Exported 6684 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION EB3 (microtubule-associated protein) labeling with near-infrared fluorescent protein miRFP670nano. ACCESSION . VERSION . KEYWORDS pEB3-miRFP670nano SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6684) AUTHORS Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV TITLE Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing. JOURNAL Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8. PUBMED 30655515 REFERENCE 2 (bases 1 to 6684) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6684 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 124..503 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 504..707 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 658..678 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 752..771 /label=T7 /note="T7 promoter, forward primer" promoter 752..770 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2126..2144) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2127..2144) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2164..2181) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2170..2394 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2440..2868 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2527..2546) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2737..2758 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2877..2897) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2882..3211 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3062..3197 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3124..3143 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3278..4072 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3332..3351) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3942..3961 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4246..4367 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4283..4302) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4337..4356 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4416..4432) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4416..4432) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4429..4451) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4440..4456 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4464..4494) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4509..4530 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4647..4664) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4818..5406) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4898..4917) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5577..6437) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6200..6219 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6438..6542) /gene="bla" /label=AmpR promoter primer_bind complement(6617..6636) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 61 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 121 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 481 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 541 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 601 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 721 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 781 cttggtacca tggccgtcaa tgtgtactcc acatctgtga ccagtgaaaa tctgagtcgc 841 catgatatgc ttgcatgggt caacgactcc ctgcacctca actataccaa gatagaacag 901 ctttgttcag gggcagccta ctgccagttc atggacatgc tcttccccgg ctgtgtgcac 961 ttgaggaaag tgaagttcca ggccaaacta gagcatgaat acatccacaa cttcaaggtg 1021 ctgcaagcag ctttcaagaa gatgggtgtt gacaaaatca ttcctgtaga gaaattagtg 1081 aaaggaaaat tccaagataa ttttgagttt attcagtggt ttaagaaatt ctttgacgca 1141 aactatgatg gaaaggatta caaccctctg ctggcgcggc agggccagga cgtagcgcca 1201 cctcctaacc caggtgatca gatcttcaac aaatccaaga aactcattgg cacagcagtt 1261 ccacagagga cgtcccccac aggcccaaaa aacatgcaga cctctggccg gctgagcaat 1321 gtggcccccc cctgcattct ccggaagaat cctccatcag cccgaaatgg cggccatgag 1381 actgatgccc aaattcttga actcaaccaa cagctggtgg acttgaagct gacagtggat 1441 gggctggaga aggaacgtga cttctacttc agcaaacttc gtgacatcga gctcatctgc 1501 caggagcatg aaagtgaaaa cagccctgtt atctcaggca tcattggcat cctctatgcc 1561 acagaggaag gattcgcacc ccctgaggac gatgagattg aagagcatca acaagaagac 1621 caggacgagt acctggatcc accggtaatg gcaaacctgg acaagatgct gaataccaca 1681 gtaacagagg tgcggcagtt cctgcaggtg gacagagtgt gcgtgttcca gtttgaggag 1741 gattatagcg gagtggtggt ggtggaggcc gtggacgata ggtggatctc catcctgaag 1801 acccaggtgc gggatagata cttcatggag acaaggggcg aggagtattc tcacggccgc 1861 taccaggcca tcgccgacat ctacaccgca aacctgacag agtgctacag ggatctgctg 1921 acacagtttc aggtgagagc aatcctggcc gtgcccatcc tgcagggcaa gaagctgtgg 1981 ggcctgttgg tggcacacca gctggcggcc cctagacagt ggcagacctg ggagatcgac 2041 tttctgaagc agcaggccgt ggtggtgggc atcgccatcc agcagagcta ggcggccgct 2101 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat 2161 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 2221 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 2281 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 2341 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg 2401 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat 2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 2641 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 2761 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa 2881 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2941 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 3001 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 3061 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 3121 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg 3181 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 3241 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 3301 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 3361 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 3421 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 3481 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 3541 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 3601 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 3661 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 3721 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 3781 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 3841 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 3901 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 3961 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 4021 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 4081 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 4141 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 4201 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 4261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 4321 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 4381 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 4441 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 4501 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 4561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 4621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 4681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 4741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 4981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 5221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 5341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 5401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 5461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 5521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 5581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 5641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 5701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 5761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 5821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 5881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 5941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 6001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 6061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 6121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 6181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 6241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 6301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 6361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 6421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 6481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 6541 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc 6601 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc 6661 ctgcttgtgt gttggaggtc gctg //