LOCUS Exported 5420 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Produces a fusion between mTurquoise2 and sReach. It can be used as a positive control for FRET (using FLIM). ACCESSION . VERSION . KEYWORDS sReach-mTurquoise2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5420) AUTHORS Mastop M, Bindels DS, Shaner NC, Postma M, Gadella TWJ Jr, Goedhart J TITLE Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2. JOURNAL Sci Rep. 2017 Sep 20;7(1):11999. doi: 10.1038/s41598-017-12212-x. PUBMED 28931898 REFERENCE 2 (bases 1 to 5420) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5420 /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 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1078..1329 /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH YLSYQSALSKDPNEKRDHMVLLERVTAAGITLGMDELYK" regulatory 1360..1369 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1366..2085 /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 2019..2040 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2208..2329 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2245..2264) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2299..2318 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2336..2791) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2473..2494) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2685..2704 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2818..2922 /gene="bla" /label=AmpR promoter promoter 2924..3281 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2947..2967) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3132..3267 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3194..3213 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3316..4110 /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(3370..3389) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3980..3999 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4298..4317) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4342..4389 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4718..5306 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5207..5226 /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ctcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactgg aacagcgtca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gagagtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atcttcggat ccaccggtcg ccaccatggt gagcaagggc 1381 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 1441 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 1501 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 1561 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 1621 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 1681 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 1741 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 1801 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac 1861 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag 1921 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 1981 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 2041 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct 2101 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 2161 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 2221 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 2281 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt 2341 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 2401 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 2461 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 2521 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 2581 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 2641 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 2701 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 2761 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc 2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc 2941 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 3001 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3121 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3181 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 3241 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga 3301 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3361 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3421 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3481 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3541 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3601 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3661 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3721 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 3781 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 3841 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 3901 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 3961 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 4021 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4081 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4141 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4201 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4261 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac 4321 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt 4381 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga 4441 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt 4501 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag 4561 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 4621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 4681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 4741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 4801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 4861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 4921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 4981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 5041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 5101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 5161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 5221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 5281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 5341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 5401 accgtattac cgccatgcat //