LOCUS Exported 3606 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Photoconverible Fluorescent Protein_ Fixative Resistant (monomeric) . ACCESSION . VERSION . KEYWORDS pRSETa_ mEos4b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3606) AUTHORS Paez-Segala MG, Sun MG, Shtengel G, Viswanathan S, Baird MA, Macklin JJ, Patel R, Allen JR, Howe ES, Piszczek G, Hess HF, Davidson MW, Wang Y, Looger LL TITLE Fixation-resistant photoactivatable fluorescent proteins for CLEM. JOURNAL Nat Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225. PUBMED 25581799 REFERENCE 2 (bases 1 to 3606) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3606 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 19..38 /label=T7 /note="T7 promoter, forward primer" promoter 19..37 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 68..90 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 110..127 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 131..163 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 167..190 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 206..889 /codon_start=1 /product="fixation-resistant truly monomeric green-to-red photoswitchable fluorescent protein (Paez-Segala et al., 2015)" /label=mEos4b /note="derived from mEos2" /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGDGTGKPYEGKQTMDLEV KEGGPLPFAFDILTTAFHYGNRVFVKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNA RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL LEGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP DNARR" primer_bind complement(952..970) /label=T7 Term /note="T7 terminator, reverse primer" terminator 966..1013 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1110..1565 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1197..1216) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1407..1428 /label=F1ori-F /note="F1 origin, forward primer" promoter 1646..1750 /gene="bla" /label=AmpR promoter rep_origin 2782..3370 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3271..3290 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3524..3541 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta 61 gaataatttt gtttaacttt aagaaggaga tatacatatg cggggttctc atcatcatca 121 tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga 181 tgacgataag gatcgatggg gatccatggt gagtgcgatt aagccagaca tgaggatcaa 241 actccgtatg gaaggcaacg taaacgggca ccactttgtg atcgacggag atggtacagg 301 caagccttat gagggaaaac agaccatgga tcttgaagtc aaagagggcg gacctctgcc 361 ttttgccttt gatatcctga ccactgcatt ccattacggc aacagggtat tcgtgaaata 421 tccagacaac atacaagact attttaagca gtcgtttcct aaggggtatt cgtgggaacg 481 aagcttgact ttcgaagacg ggggcatttg caatgccaga aacgacataa caatggaagg 541 ggacactttc tataataaag ttcgatttta tggtaccaac tttcccgcca atggtccagt 601 tatgcagaag aagacgctga aatgggagcc ctccactgag aaaatgtatg tgcgtgatgg 661 agtgctgacg ggtgatattg agatggcttt gttgcttgaa ggaaatgccc attaccgatg 721 tgacttcaga actacttaca aagctaagga gaagggtgtc aagttaccag gcgcccactt 781 tgtggaccac gccattgaga ttttaagcca tgacaaagat tacaacaagg ttaagctgta 841 tgagcatgct gttgctcatt ctggattgcc tgacaatgcc agacgataag aattcgaagc 901 ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc 961 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag 1021 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca 1081 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc 1141 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 1201 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 1261 tcgggggctc cctttagggt tccgatttag agctttacgg cacctcgacc gcaaaaaact 1321 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 1381 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 1441 ccctatcgcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 1501 aaaaaatgag ctgatttaac aaatatttaa cgcgaatttt aacaaaatat taacgtttac 1561 aatttcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac 1621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 1681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 1741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 1801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 1861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 1921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtgatac 1981 actattatcc cgtattgacg ccgggcaaga gcaactcggt cgccccccgc ggtattctca 2041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 2101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 2161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 2221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagagtga 2281 caccacgatg cctgtagcaa tgccaacaac gttgcgcaaa ctattaactg gcgaactact 2341 tactctagct tcccggcaac aattaataga ctgaatggag gcggataaag ttgcaggacc 2401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 2461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagcgct cccgtatcgt 2521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 2581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 2641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 2701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 2761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 2821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 2881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 2941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 3001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 3061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 3121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 3181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 3241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 3301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 3361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctgggctttt gctggccttt 3421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 3481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 3541 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 3601 atgcag //