LOCUS Exported 3580 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION bacterial expression of Sirius, a pH-insensitive and photostable ultramarine fluorescent protein. ACCESSION . VERSION . KEYWORDS Sirius/pRSETB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3580) AUTHORS Tomosugi W, Matsuda T, Tani T, Nemoto T, Kotera I, Saito K, Horikawa K, Nagai T TITLE An ultramarine fluorescent protein with increased photostability and pH insensitivity. JOURNAL Nat Methods. 2009 May;6(5):351-3. doi: 10.1038/nmeth.1317. Epub 2009 Apr 6. PUBMED 19349978 REFERENCE 2 (bases 1 to 3580) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3580 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 70..92 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 112..129 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 133..165 /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 169..192 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" primer_bind complement(244..265) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(505..524) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 664..915 /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKAHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH YLSVQSKLSKDPNEKRDHMVLLESVTAAGITLGMDELYK" primer_bind complement(981..999) /label=T7 Term /note="T7 terminator, reverse primer" terminator 995..1042 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1139..1594 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1226..1245) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1436..1457 /label=F1ori-F /note="F1 origin, forward primer" promoter 1620..1724 /gene="bla" /label=AmpR promoter CDS 1725..2585 /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(1943..1962) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2756..3344 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3245..3264 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3498..3515 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat 121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac 181 gatgacgata aggatccgat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 241 atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc 301 gagggcgatg ccacctacgg caagctgacc ctgaagctca tctgcaccac cggcaagctg 361 cccgtgccct ggcccaccct cgtgaccacc ctgcaattcg gcgtgctgtg cttcgcccgc 421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 481 caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 601 ggcaacatcc tggggcacaa gctggagtac aacgggataa gctcaaacgt atatatcacc 661 gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac 721 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 781 ctgctgcccg acaaccacta cctgagcgtc cagtccaagc tgagcaaaga ccccaacgag 841 aagcgcgatc acatggtcct gctggagtcc gtgaccgccg ccgggatcac tctcggcatg 901 gacgagctgt acaagtaaga attcgaagct tgatccggct gctaacaaag cccgaaagga 961 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa 1021 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc 1081 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac 1141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 1201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 1261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 1321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 1381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 1441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 1501 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 1561 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc 1621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 2641 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 2701 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 2761 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 2821 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 2881 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 2941 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 3001 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 3061 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 3121 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 3181 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 3241 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 3301 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 3361 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 3421 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 3481 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 3541 aaccgcctct ccccgcgcgt tggccgattc attaatgcag //