LOCUS Exported 3618 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of mCyRFP1. ACCESSION . VERSION . KEYWORDS pNCS-mCyRFP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3618) AUTHORS Laviv T, Kim BB, Chu J, Lam AJ, Lin MZ, Yasuda R TITLE Simultaneous dual-color fluorescence lifetime imaging with novel red-shifted fluorescent proteins. JOURNAL Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046. PUBMED 27798609 REFERENCE 2 (bases 1 to 3618) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3618 /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" CDS 164..181 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 185..217 /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 221..244 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 251..952 /codon_start=1 /product="monomeric cyan-excitable red fluorescent protein (Laviv et al., 2016)" /label=mCyRFP1 /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTA RIKVVEGGPLPFAFDILATMFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG VLTATQDTSLQDGELIYNVKLRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD KKLKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY SNLGGGMDELYK" primer_bind complement(1019..1037) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1033..1080 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1177..1632 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1264..1283) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1474..1495 /label=F1ori-F /note="F1 origin, forward primer" promoter 1658..1762 /gene="bla" /label=AmpR promoter CDS 1763..2623 /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(1981..2000) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2794..3382 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3283..3302 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3536..3553 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg 121 ttttgtttaa ctttaagaag gggatataca tatgcggggt tctcatcatc atcatcatca 181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga 241 taaggatccg atggtgagca agggcgagga gctgatcaag gagaacatga gaagcaagct 301 gtacctggaa ggcagcgtga acggccacca gttcaagtgc acccacgaag gggagggcaa 361 gccctacgag ggcaagcaga ccgcgaggat caaggtggtg gagggaggcc ccctgccgtt 421 cgcattcgac atcctggcca ccatgtttat gtacgggagc aaggtgttca tcaagtaccc 481 cgccgacctc cccgattatt ttaagcagtc cttccctgag ggcttcacat gggagagagt 541 catggtgttc gaagacgggg gcgtgctgac cgccacccag gacaccagcc tccaggacgg 601 cgagctcatc tacaacgtca agctcagagg ggtgaacttc ccagccaacg gccccgtgat 661 gcagaagaaa acactgggct gggagcccag caccgagacc atgtaccccg ctgacggcgg 721 cctggaaggc agatgcgaca agaagctgaa gctcgtgggc gggggccacc tgcacgtcaa 781 cttcaagacc acatacaagt ccaagaaacc cgtgaagatg cccggcgtcc actacgtgga 841 ccgcagactg gaaagaatca aggaggccga caacgagacc tacgtcgagc agtacgagca 901 cgctgtggcc agatactcca acctgggcgg aggcatggac gagctgtaca agtaatgaat 961 tcgaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 1021 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 1081 ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc accgatcgcc 1141 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa 1201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 1261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 1321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 1381 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 1441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 1501 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 1561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 1621 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 1681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 1741 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 1801 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 1861 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 1921 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 1981 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 2041 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 2101 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 2161 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 2221 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 2281 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 2341 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 2401 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 2461 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 2521 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 2581 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 2641 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 2701 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 2761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 2821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 2881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 2941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 3001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 3121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 3181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 3241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 3301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 3361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 3421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 3481 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 3541 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 3601 gccgattcat taatgcag //