LOCUS Exported 5929 bp ds-DNA circular SYN 04-OCT-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS nuclear EKAR (EGFP-mRFP) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5929) AUTHORS Harvey CD, Ehrhardt AG, Cellurale C, Zhong H, Yasuda R, Davis RJ, Svoboda K. TITLE A genetically encoded fluorescent sensor of ERK activity JOURNAL PNAS 2008 Dec 9;105(49):19264-9 PUBMED 19033456 REFERENCE 2 (bases 1 to 5929) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 4, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5929 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 138..517 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 518..721 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 672..692 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 718..742 /label=LNCX /note="Human CMV promoter, forward primer" intron 857..989 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 972..991 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 1034..1052 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1058..1732 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(1188..1206) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1456..1475 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1650..1673 /label=DsRed1-C /note="DsRed1, forward primer" CDS 2300..3019 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2345..2366) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2606..2625) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2953..2974 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3043..3164 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3054..3073) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3108..3127 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin 3345..3800 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3432..3451) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3642..3663 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3867..3886) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3986..4008 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4046..4064) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4132..4236 /gene="bla" /label=AmpR promoter CDS 4237..5097 /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(4455..4474) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5268..5856 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5757..5776 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat 781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc 841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa 901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac 961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta 1021 aggctagagt acttaatacg actcactata ggctagcatg gcctcctccg aggacgtcat 1081 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga 1141 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt 1201 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg 1261 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc 1321 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac 1381 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa 1441 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga 1501 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa 1561 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca 1621 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta 1681 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg ccgagctcaa 1741 gcttatggcg gacgaggaga agctgccgcc cggctgggag aagcgcatga gccgcagctc 1801 aggccgagtg tactacttca accacatcac taacgccagc cagtgggagc ggcccagcgg 1861 caacagcagc agtggtggca aaaacgggca gggggagcct gccaggaagc ttcgaattct 1921 gcagtcgact agggagcaag ggcttgtgtc tagaggcgga ggtggtggag gaggtggagg 1981 tggtggaggt ggaggtggag gtggtggcgg tggtggagga ggaggagcta gaggcggagg 2041 tggtggaggr ggkggaggtg gtggaggtgg aggtggaggt ggtggsggtg gtggaggagg 2101 aggagctagm ggcggaggtg gtggaggagg tggaggtggt ggaggtggag gtggaggtgg 2161 tggcggtggt ggaggaggag gaatggctga ccaactgact gaagagtggt cgacggtacc 2221 gcgggatatc cctgacgtac cacgtacacc agtaggaaag gcaaagctgt cattccaatt 2281 cccgcgggcc cgggatccca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 2341 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 2401 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 2461 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 2521 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 2581 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 2641 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 2701 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 2761 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 2821 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 2881 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga 2941 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 3001 ggacgagctg tacaagtaag cggccgcttc gagcagacat gataagatac attgatgagt 3061 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 3121 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 3181 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc 3241 tctacaaatg tggtaaaatc gataaggatc cgggctggcg taatagcgaa gaggcccgca 3301 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg 3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 3781 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 3841 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 3901 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc 3961 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca 4021 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc 4081 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 4141 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 4201 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 4261 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 4321 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 4381 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 4441 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 4501 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 4561 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 4621 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 4681 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 4741 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 4801 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 4861 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 4921 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 4981 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 5041 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 5101 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 5161 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 5221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 5281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 5341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 5401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta 5461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 5521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 5581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 5641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 5701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 5761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 5821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 5881 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatct //