LOCUS Exported 8018 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses iPOLYMER component in mammalian cells. ACCESSION . VERSION . KEYWORDS pMT2 TIA1 RRM-CFP-FRBx5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8018) AUTHORS Nakamura H, Lee AA, Afshar AS, Watanabe S, Rho E, Razavi S, Suarez A, Lin YC, Tanigawa M, Huang B, DeRose R, Bobb D, Hong W, Gabelli SB, Goutsias J, Inoue T TITLE Intracellular production of hydrogels and synthetic RNA granules by multivalent molecular interactions. JOURNAL Nat Mater. 2017 Nov 6. pii: nmat5006. doi: 10.1038/nmat5006. PUBMED 29115293 REFERENCE 2 (bases 1 to 8018) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8018 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(7..336) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 21..156 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(75..94) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 321..341 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" intron 805..1034 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 1017..1036 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1062..1071 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 1731..1740 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1737..2453 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1782..1803) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2043..2062) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2390..2411 /label=EGFP-C /note="EGFP, forward primer" CDS 2466..2744 /codon_start=1 /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /label=FRB* /note="binds synthetic ligands such as AP21967" /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" CDS 2835..3113 /codon_start=1 /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /label=FRB* /note="binds synthetic ligands such as AP21967" /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" CDS 3156..3434 /codon_start=1 /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /label=FRB* /note="binds synthetic ligands such as AP21967" /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" CDS 3477..3755 /codon_start=1 /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /label=FRB* /note="binds synthetic ligands such as AP21967" /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" CDS 3798..4076 /codon_start=1 /product="T2098L mutant of FKBP-rapamycin binding domain of human FRAP" /label=FRB* /note="binds synthetic ligands such as AP21967" /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK" polyA_signal 4969..5103 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5006..5025) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5060..5079 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5630..5647) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5801..6389) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5881..5900) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6560..7420) /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 7183..7202 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7421..7525) /gene="bla" /label=AmpR promoter primer_bind 7593..7611 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7649..7671) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7771..7790 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7984..8006 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7998..8015 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7999..8015 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc 61 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg 121 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat 181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga 241 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 301 ggagcctggg gactttccac accctaactg acacacattc cacaggatcc ggtcgcgcga 361 atttcgagcg gtgttccgcg gtcctcctcg tatagaaact cggaccactc tgagacgaag 421 gctcgcgtcc aggccagcac gaaggaggct aagtgggagg ggtagcggtc gttgtccact 481 agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc cctcttcggc atcaaggaag 541 gtgattggtt tataggtgta ggccacgtga ccgggtgttc ctgaaggggg gctataaaag 601 ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt 661 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc 721 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga 781 tcggaaaacc tctcgactgt tggggtgagt actccctctc aaaagcgggc atgacttctg 841 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga 901 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct 961 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc 1021 ctttctctcc acaggtgtcc actcccaggt ccaactgcag ggccaccatg gaggacgaga 1081 tgcccaagac tctatacgtc ggtaaccttt ccagagatgt gacagaagct ctaattctgc 1141 aactctttag ccagattgga ccttgtaaaa actgcaaaat gattatggat acagctggaa 1201 atgatcccta ttgttttgtg gagtttcatg agcatcgtca tgcagctgca gcattagctg 1261 ctatgaatgg acggaagata atgggtaagg aagtcaaagt gaattgggca acaaccccta 1321 gcagtcaaaa gaaagataca agcaatcatt tccatgtctt tgttggtgat ctcagcccag 1381 aaattacaac tgaagatata aaagctgctt ttgcaccatt tggaagaata tcagatgccc 1441 gagtggtaaa agacatggca acaggaaagt ctaagggata tggctttgtc tcctttttca 1501 acaaatggga tgctgaaaac gccattcaac agatgggtgg ccagtggctt ggtggaagac 1561 aaatcagaac taactgggca acccgaaagc ctcccgctcc aaagagtaca tatgagtcaa 1621 ataccaaaca gctatcatat gatgaggttg taaatcagtc tagtccaagc aactgtactg 1681 tatactgtgg aggtgttact tctgggctaa cagaacaact aggaccggtc gccaccatgg 1741 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 1801 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 1861 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 1921 tgaccaccct gacctggggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 1981 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 2041 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 2101 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 2161 tggagtacaa ctacatcagc cacaacgtct atatcaccgc cgacaagcag aagaacggca 2221 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 2281 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 2341 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 2401 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggagcaa 2461 gtggaatcct ctggcatgag atgtggcatg aaggcctgga agaggcatct cgtttgtact 2521 ttggggaaag gaacgtgaaa ggcatgtttg aggtgctgga gcccttgcat gctatgatgg 2581 aacggggccc ccagactctg aaggaaacat cctttaatca ggcctatggt cgagatttaa 2641 tggaggccca agagtggtgc aggaagtaca tgaaatcagg gaatgtcaag gacctcctcc 2701 aagcctggga cctctattat catgtgttcc gacgaatctc aaagagtgct ggtggtagtg 2761 ctggtggtag tgctggtggt actggtggtc ctcgagctag tgctggtggt agtgctggtg 2821 gtagtgctgg tggtatcctc tggcatgaga tgtggcatga aggcctggaa gaggcatctc 2881 gtttgtactt tggggaaagg aacgtgaaag gcatgtttga ggtgctggag cccttgcatg 2941 ctatgatgga acggggcccc cagactctga aggaaacatc ctttaatcag gcctatggtc 3001 gagatttaat ggaggcccaa gagtggtgca ggaagtacat gaaatcaggg aatgtcaagg 3061 acctcctcca agcctgggac ctctattatc atgtgttccg acgaatctca aagaagctta 3121 gtgctggtgg tagtgctggt ggtagtgctg gtggtatcct ctggcatgag atgtggcatg 3181 aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg 3241 aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat 3301 cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca 3361 tgaaatcagg gaatgtcaag gacctcctcc aagcctggga cctctattat catgtgttcc 3421 gacgaatctc aaaggaattc agtgctggtg gtagtgctgg tggtagtgct ggtggtatcc 3481 tctggcatga gatgtggcat gaaggcctgg aagaggcatc tcgtttgtac tttggggaaa 3541 ggaacgtgaa aggcatgttt gaggtgctgg agcccttgca tgctatgatg gaacggggcc 3601 cccagactct gaaggaaaca tcctttaatc aggcctatgg tcgagattta atggaggccc 3661 aagagtggtg caggaagtac atgaaatcag ggaatgtcaa ggacctcctc caagcctggg 3721 acctctatta tcatgtgttc cgacgaatct caaaggtcga cagtgctggt ggtagtgctg 3781 gtggtagtgc tggtggtatc ctctggcatg agatgtggca tgaaggcctg gaagaggcat 3841 ctcgtttgta ctttggggaa aggaacgtga aaggcatgtt tgaggtgctg gagcccttgc 3901 atgctatgat ggaacggggc ccccagactc tgaaggaaac atcctttaat caggcctatg 3961 gtcgagattt aatggaggcc caagagtggt gcaggaagta catgaaatca gggaatgtca 4021 aggacctcct ccaagcctgg gacctctatt atcatgtgtt ccgacgaatc tcaaagccgc 4081 gggcccggga tccaccggat ctagataact gacaattcgg gggggggggg gggggggaca 4141 gctcagggct gcgatttcgc gccaaacttg acggcaatcc tagcgtgaag gctggtagga 4201 ttttatcccc gctgccatca tggttcgacc attgaactgc atcgtcgccg tgtcccaaaa 4261 tatggggatt ggcaagaacg gagacctacc ctggcctccg ctcaggaacg agttcaagta 4321 cttccaaaga atgaccacaa cctcttcagt ggaaggtaaa cagaatctgg tgattatggg 4381 taggaaaacc tggttctcca ttcctgagaa gaatcgacct ttaaaggaca gaattaatat 4441 agttctcagt agagaactca aagaaccacc acgaggagct cattttcttg ccaaaagttt 4501 ggatgatgcc ttaagactta ttgaacaacc ggaattggca agtaaagtag acatggttgg 4561 atagtcggag gcagttctgt ttaccaggaa gccatgaatc aaccaggcca cctcagactc 4621 tttgtgacaa ggatcatgca ggaatttgaa agtgacacgt ttttcccaga aattgatttg 4681 gggaaatata aacttctccc agaataccca ggcgtcctct ctgaggtcca ggaggaaaaa 4741 ggcatcaagt ataagtttga agtctacgag aagaaagact aacaggaaga tgctttcaag 4801 ttctctgctc ccctcctaaa gctatgcatt tttataagac catgggactt ttgctggctt 4861 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 4921 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 4981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 5041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 5101 atccccggcc aacggtctgg tgacccggct gcgagagctc ggtgtacctg agacgcgagt 5161 aagcccttga gtcaaagacg tagtcgttgc aagtccgcac caggtactga tatcccacca 5221 aaaagtgcgg cggcggctgg cggtagaggg gccagcgtag ggtggccggg gctccggggg 5281 cgaggtcttc caacataagg cgatgatcat cgatgctaga ccgtgcaaaa ggagagcctg 5341 taagcgggca ctcttccgtg gtctggtgga taaattcgca agggtatcat ggcggacgac 5401 cggggttcga accccggatc cggccgtccg ccgtgatcca tgcggttacc gcccgcgtgt 5461 cgaacccagg tgtgcgacgt cagacaacgg gggagcgctc cttttggctt ccttccaggc 5521 gcggcggctg ctgcgctagc ttttttggcg agctcgaatt aattctgcat taatgaatcg 5581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 7561 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 7621 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 7681 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 7741 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 7801 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 7861 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 7921 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 7981 tttcccagtc acgacgttgt aaaacgacgg ccagtgcc //