LOCUS Exported 4793 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the nucleus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS 3xnls-mTurquoise2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4793) AUTHORS Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J TITLE Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells JOURNAL bioRxiv 160374 REFERENCE 2 (bases 1 to 4793) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4793 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 113..132 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 372..393 /label=EGFP-N /note="EGFP, reverse primer" regulatory 435..444 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(457..477) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(481..501) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(505..525) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter complement(609..812) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(638..658) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 813..1116 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(1291..1879) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1371..1390) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 2208..2255 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 2280..2299 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(2487..3281) /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(2598..2617) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3208..3227 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(3316..3673) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3330..3465 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3384..3403) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 3630..3650 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(3675..3779) /gene="bla" /label=AmpR promoter rep_origin 3806..4261 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3893..3912) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4103..4124 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 4268..4389 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4279..4298) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4333..4352 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(4512..438) /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4557..4578) /label=EGFP-C /note="EGFP, forward primer" ORIGIN 1 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt 61 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta 121 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga 181 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgcccca 241 ggacagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa 301 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt 361 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc 421 ctcgcccttg ctcaccatgg tggcgaccgg tgatcccact ttccgtttct ttttgggatc 481 gaccttacgc tttttcttcg ggtctacctt tctcttcttt tttggatctg atcccatggt 541 gggaattcga agcttgagct cgagatctga gtccggtagc gctagcggat ctgacggttc 601 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc 661 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa 721 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca 781 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag 841 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc 901 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact 961 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt 1021 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt 1081 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga 1141 actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc 1201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 1321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 1381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 1441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 1501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 1561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 1621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 1681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 1741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga 2041 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact 2101 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg 2161 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc 2221 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat 2281 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag 2341 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct 2401 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc 2461 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg 2521 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg 2581 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc 2641 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc 2701 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg 2761 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga 2821 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg 2881 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg 2941 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc 3001 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg 3061 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg 3121 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg 3181 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa 3241 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct 3301 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg 3361 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc 3421 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg 3481 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga 3541 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct 3601 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt 3661 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt 3721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 3781 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt 3841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 4201 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt 4261 tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 4321 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 4381 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 4441 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctagag 4501 tcgcggccgc tttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg 4561 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg 4621 gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc 4681 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg 4741 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtcgct aaa //