LOCUS Exported 3550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION bacterial expression of an EosFP variant, mEos2 (a photoconvertible fluorescent protein). ACCESSION . VERSION . KEYWORDS pRSETa mEos2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3550) AUTHORS McKinney SA, Murphy CS, Hazelwood KL, Davidson MW, Looger LL TITLE A bright and photostable photoconvertible fluorescent protein. JOURNAL Nat Methods. 2009 Feb . 6(2):131-3. PUBMED 19169260 REFERENCE 2 (bases 1 to 3550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3550 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(650..668) /label=T7 Term /note="T7 terminator, reverse primer" terminator 664..711 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 808..1263 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(895..914) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1105..1126 /label=F1ori-F /note="F1 origin, forward primer" promoter 1289..1393 /gene="bla" /label=AmpR promoter CDS 1394..2254 /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 complement(1612..1631) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2425..3013 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2914..2933 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3167..3184 /label=L4440 /note="L4440 vector, forward primer" primer_bind 3269..3288 /label=T7 /note="T7 promoter, forward primer" promoter 3269..3287 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 3319..3341 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 3361..3378 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 3382..3414 /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 3418..3441 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 3457..587 /codon_start=1 /product="monomeric variant of green-to-red photoswitchable fluorescent protein EosFP (McKinney et al., 2009)" /label=mEos2 /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD NARR" ORIGIN 1 agccttttga gggaaaacag agtatggatc ttgaagtcaa agagggcgga cctctgcctt 61 ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc gccaaatatc 121 cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa 181 gcttgacttt cgaagacggg ggcatttgca ttgccagaaa cgacataaca atggaagggg 241 acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta 301 tgcagaagaa gacgctgaaa tgggagccct ccactgagaa aatgtatgtg cgtgatggag 361 tgctgacggg tgatattcat atggctttgt tgcttgaagg aaatgcccat taccgatgtg 421 acttcagaac tacttacaaa gctaaggaga agggtgtcaa gttaccaggc taccactttg 481 tggaccactg cattgagatt ttaagccatg acaaagatta caacaaggtt aagctgtatg 541 agcatgctgt tgctcattct ggattgcctg acaatgccag acgataagaa ttcgaagctt 601 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 661 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 721 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 781 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 1021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 1081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 1141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 1201 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 1261 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 2101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 3061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 3121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 3181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 3241 ttaatgcagg atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg 3301 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct 3361 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat 3421 ctgtacgacg atgacgataa ggatcgatgg ggatccatga gtgcgattaa gccagacatg 3481 aagatcaaac tccgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat 3541 ggtacaggca //