LOCUS Exported 4778 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of the cytoplasm, but not the nucleus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS pmTurquoise2-NES SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4778) AUTHORS Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A TITLE Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. JOURNAL Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. PUBMED 22434194 REFERENCE 2 (bases 1 to 4778) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4778 /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" misc_feature 591..671 /label=MCS /note="multiple cloning site" regulatory 673..682 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 679..1392 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELY" primer_bind complement(724..745) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(985..1004) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1332..1353 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1566..1687 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1603..1622) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1657..1676 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1694..2149) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1831..1852) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2043..2062 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2176..2280 /gene="bla" /label=AmpR promoter promoter 2282..2639 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2305..2325) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2490..2625 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2552..2571 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2674..3468 /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(2728..2747) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3338..3357 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3656..3675) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3700..3747 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4076..4664 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4565..4584 /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 attctgcagt cgacggtacc gcgggcccgg 661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 841 cccgtgccct ggcccaccct cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc 901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 1081 ggcaacatcc tggggcacaa gctggagtac aactacttta gcgacaacgt ctatatcacc 1141 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac 1201 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 1261 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag 1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 1381 gacgagctgt acagtgagct gcagaacaag ctggaagagt tggatctgga ctcgtacaag 1441 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg 1501 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 1561 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 1621 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 1681 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt 1741 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 1801 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 1861 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 1921 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 1981 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 2041 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 2101 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact 2161 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 2221 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 2281 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 2341 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 2401 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2461 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 2521 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 2581 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag 2641 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 2701 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 2761 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 2821 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg 2881 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 2941 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3001 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3061 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3121 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3181 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc 3241 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 3301 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 3361 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 3421 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 3481 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 3541 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 3601 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa 3661 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata 3721 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact 3781 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc 3841 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag 3901 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt 3961 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 4021 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 4081 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 4141 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 4201 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 4261 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 4321 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 4381 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 4441 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 4501 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 4561 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 4621 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 4681 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 4741 cccctgattc tgtggataac cgtattaccg ccatgcat //