WO 22F323WO JP 2022-033316 2022-03-04 JAPAN SCIENCE AND TECHNOLOGY AGENCY Capped RNA and method for producing thereof, protein production device and method for producing protein 64 20 RNA PAT misc_feature 1..20 note RNA1 misc_feature 1 note 5' is monophosphate source 1..20 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgcc 21 RNA PAT misc_feature 1..21 note RNA2 misc_feature 1 note 5' is monophosphate misc_feature 1^2 note C4 linker source 1..21 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgcc 20 RNA PAT misc_feature 1..20 note RNA3 misc_feature 1 note 5' is monophosphate misc_feature 1 note C4 linker source 1..20 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgcc 20 RNA PAT misc_feature 1..20 note RNA4 misc_feature 20 note 3' is monophosphate misc_feature 20 note C4 linker source 1..20 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgcc 20 RNA PAT misc_feature 1..20 note RNA5 misc_feature 1 note 5' is monophosphate source 1..20 mol_type other RNA organism synthetic construct cgccgtggaactatctccta 50 RNA PAT misc_feature 1..50 note RNA6 misc_feature 1 note 5' is monophosphate source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note RNA7 misc_feature 1 note 5' is monophosphate misc_feature 1^2 note C4 linker source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note RNA8 misc_feature 1 note 5' is monophosphate misc_feature 1 note C4 linker source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note RNA9 misc_feature 1 note 5' is monophosphate modified_base 1 mod_base gm source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note RNA10 misc_feature 1 note 5' is monophosphate modified_base 1 mod_base gm misc_feature 1 note C4 linker source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note RNA11 misc_feature 1 note 5' is monophosphate misc_feature 1..3 note LNA misc_feature 18..20 note LNA source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note RNA12 misc_feature 1 note 5' is monophosphate misc_feature 1 note C4 linker misc_feature 1..3 note LNA misc_feature 18..20 note LNA source 1..50 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 21 RNA PAT misc_feature 1..21 note Capped RNA1 (5' capp probe in Fig.3) modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..21 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgcc 21 RNA PAT misc_feature 1..21 note Capped RNA2(5' capp probe in Fig.3) modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 2^3 note C4 linker source 1..21 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgcc 21 RNA PAT misc_feature 1..21 note Capped RNA3 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 1^2 note C4 linker source 1..21 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgcc 21 RNA PAT misc_feature 1..21 note Capped RNA4 (3' capp probe in Fig.3) misc_feature 20^21 note triphosphate linkage misc_feature 20^21 note C4 linker modified_base 21 mod_base m7g source 1..21 mol_type other RNA organism synthetic construct gcgtcaccttaatatgcgccg 21 RNA PAT misc_feature 1..21 note Capped RNA5 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..21 mol_type other RNA organism synthetic construct gcgccgtggaactatctccta 51 RNA PAT misc_feature 1..51 note Capped RNA6 modified_base 1 mod_base m7g source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 52 RNA PAT misc_feature 1..52 note Capped RNA7 (5' capp probe with poly A in Fig.3) modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 2^3 note C4 linker source 1..52 mol_type other RNA organism synthetic construct gggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note Capped RNA8 (5' capp probe with poly A in Fig.3) modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 1^2 note C4 linker source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note Capped RNA9 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage modified_base 2 mod_base gm source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note Capped RNA10 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 1^2 note C4 linker modified_base 2 mod_base gm source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note Capped RNA11 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 2..4 note LNA misc_feature 18..20 note LNA source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT misc_feature 1..51 note Capped RNA12 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage misc_feature 1^2 note C4 linker misc_feature 2..4 note LNA misc_feature 18..20 note LNA source 1..51 mol_type other RNA organism synthetic construct ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 48 DNA PAT misc_feature 1..48 note primer_Fw source 1..48 mol_type other DNA organism synthetic construct cccggatcctaatacgactcactataggtcttcacactcgaagatttc 51 DNA PAT misc_feature 1..51 note primer_Rev source 1..51 mol_type other DNA organism synthetic construct ttttttttttttttttttttttttttttttctagaattacgccagaatgcg 30 DNA PAT misc_feature 1..30 note Splint_1 source 1..30 mol_type other DNA organism synthetic construct accttaatatgcgccttttttttttttttt 30 DNA PAT misc_feature 1..30 note Splint_2 source 1..30 mol_type other DNA organism synthetic construct tcgagtgtgaagaccatggtggctttaccc 107 RNA PAT misc_feature 1..107 note blanch RNA modified_base 1 mod_base gm misc_feature 1^2 note The 2'-methylguanosine(Gm) and the adenosine(A) are linked by abranching linker. The number of Gm's correspond to the number ofthe branches of the linker. Cap structure can be introduced tothe each 5' end of the Gm. source 1..107 mol_type other RNA organism synthetic construct ggagccaccatggactacaaggacgacgacgataagatcatcgactataaagacgacgacgataaacaccaccaccaccaccactgaaaaaaaaaaaaaaaaaaaaa 50 RNA PAT misc_feature 1..50 note bicistronic mRNA source 1..50 mol_type other RNA organism synthetic construct gacagtctgctcgaagcggccgctctagaagcgtcaccttaatatgcgcc 650 RNA PAT misc_feature 1..650 note RNA coding nanoluciferase source 1..650 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagcgaccctgcagcgacccgcttaaaagcttggcaatccggtactgttggtaaagccaccatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 676 DNA PAT misc_feature 1..676 note DNA templete of nanoluciferase coding RNA source 1..676 mol_type other DNA organism synthetic construct cccggatcctaatacgactcactataggcgcatattaaggtgacgcgtgtggcctcgaacaccgagcgaccctgcagcgacccgcttaaaagcttggcaatccggtactgttggtaaagccaccatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagtttttttttttttttttttttttttttttt 54 RNA PAT misc_feature 1..54 note Synthesis of branched RNA-1 misc_feature 14^15 note The guanosine(G) and the uridine(U) are linked by a branchinglinker. GmGmAGCG is linked to the branched chain that branchesoff from the main chain. ("Gm" indicates 2'OMe guanosine). A capstructure can be attached to the 5' end of the Gm. source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 2489 RNA PAT misc_feature 1..2489 note bicistronic RNA coding firefly-luciferase and nano-luciferase modified_base 1 mod_base m7g source 1..2489 mol_type other RNA organism synthetic construct gggcgaattcgagcggcattccggtactgttggttttgccaccatggactacaaggacgacgacgacaagatcatcgactataaagacgacgacgataaaggtggcgactataaggacgacgacgacaaagccaccatggaagacgccaaaaacataaagaaaggcccggcgccattctatccgctggaagatggaaccgctggagagcaactgcataaggctatgaagagatacgccctggttcctggaacaattgcttttacagatgcacatatcgaggtggacatcacttacgctgagtacttcgaaatgtccgttcggttggcagaagctatgaaacgatatgggctgaatacaaatcacagaatcgtcgtatgcagtgaaaactctcttcaattctttatgccggtgttgggcgcgttatttatcggagttgcagttgcgcccgcgaacgacatttataatgaacgtgaattgctcaacagtatgggcatttcgcagcctaccgtggtgttcgtttccaaaaaggggttgcaaaaaattttgaacgtgcaaaaaaagctcccaatcatccaaaaaattattatcatggattctaaaacggattaccagggatttcagtcgatgtacacgttcgtcacatctcatctacctcccggttttaatgaatacgattttgtgccagagtccttcgatagggacaagacaattgcactgatcatgaactcctctggatctactggtctgcctaaaggtgtcgctctgcctcatagaactgcctgcgtgagattctcgcatgccagagatcctatttttggcaatcaaatcattccggatactgcgattttaagtgttgttccattccatcacggttttggaatgtttactacactcggatatttgatatgtggatttcgagtcgtcttaatgtatagatttgaagaagagctgtttctgaggagccttcaggattacaagattcaaagtgcgctgctggtgccaaccctattctccttcttcgccaaaagcactctgattgacaaatacgatttatctaatttacacgaaattgcttctggtggcgctcccctctctaaggaagtcggggaagcggttgccaagaggttccatctgccaggtatcaggcaaggatatgggctcactgagactacatcagctattctgattacacccgagggggatgataaaccgggcgcggtcggtaaagttgttccattttttgaagcgaaggttgtggatctggataccgggaaaacgctgggcgttaatcaaagaggcgaactgtgtgtgagaggtcctatgattatgtccggttatgtaaacaatccggaagcgaccaacgccttgattgacaaggatggatggctacattctggagacatagcttactgggacgaagacgaacacttcttcatcgttgaccgcctgaagtctctgattaagtacaaaggctatcaggtggctcccgctgaattggaatccatcttgctccaacaccccaacatcttcgacgcaggtgtcgcaggtcttcccgacgatgacgccggtgaacttcccgccgccgttgttgttttggagcacggaaagacgatgacggaaaaagagatcgtggattacgtcgccagtcaagtaacaaccgcgaaaaagttgcgcggaggagttgtgtttgtggacgaagtaccgaaaggtcttaccggaaaactcgacgcaagaaaaatcagagagatcctcataaaggccaagaagggcggaaagatcgccgtgtaattctagagtcggggcggccggccgcttcgagcagacacgataagatacagggcgcatattaaggtgacgcgtgtggcctcgaacaccgagcgaccctgcagcgacccgcttaaaagcttggcaatccggtactgttggtaaagccaccatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 24 RNA PAT misc_feature 1..24 note RNA5 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..24 mol_type other RNA organism synthetic construct ggaacggttacagcaacgctgctg 24 RNA PAT misc_feature 1..24 note RNA6 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..24 mol_type other RNA organism synthetic construct gaccgctttgctggaatcgaggaa 24 RNA PAT misc_feature 1..24 note RNA7 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..24 mol_type other RNA organism synthetic construct ggataggtgtctttgtcggttaca 24 RNA PAT misc_feature 1..24 note RNA8 modified_base 1 mod_base m7g misc_feature 1^2 note triphosphate linkage source 1..24 mol_type other RNA organism synthetic construct ggatttgcggtgctttgctggaat 1338 DNA PAT misc_feature 1..1338 note mRNA coding Renilla luciferase source 1..1338 mol_type other DNA organism synthetic construct ggtttctactttgcccgcccacagatgtagttttctctgcgcgtgtgcgttttccctcctccccgccctcagggtccacggccaccatggcgtattaggggcagcagtgcctgcggcagcattggcctttgcagcggcggcagcagcaccaggctctgcagcggcaacccccagcggcttaagccatggcgcttctcacggcattcagcagcagcgttgctgtaaccgacaaagacaccttcgaattaagcacattcctcgattccagcaaagcaccgcaacatgaccgaaatgagcttcctgagcagcgaggtgttggtgggggacttgatgtcccccttcgaccagtcgggtttgggggctgaggcttccaaggtgtacgaccccgagcaacgcaaacgcatgatcactgggcctcagtggtgggctcgctgcaagcaaatgaacgtgctggactccttcatcaactactatgattccgagaagcacgccgagaacgccgtgatttttctgcatggtaacgctgcctccagctacctgtggaggcacgtcgtgcctcacatcgagcccgtggctagatgcatcatccctgatctgatcggaatgggtaagtccggcaagagcgggaatggctcatatcgcctcctggatcactacaagtacctcaccgcttggttcgagctgctgaaccttccaaagaaaatcatctttgtgggccacgactggggggcttgtctggcctttcactactcctacgagcaccaagacaagatcaaggccatcgtccatgctgagagtgtcgtggacgtgatcgagtcctgggacgagtggcctgacatcgaggaggatatcgccctgatcaagagcgaagagggcgagaaaatggtgcttgagaataacttcttcgtcgagaccatgctcccaagcaagatcatgcggaaactggagcctgaggagttcgctgcctacctggagccattcaaggagaagggcgaggttagacggcctaccctctcctggcctcgcgagatccctctcgttaagggaggcaagcccgacgtcgtccagattgtccgcaactacaacgcctaccttcgggccagcgacgatctgcctaagatgttcatcgagtccgaccctgggttcttttccaacgctattgtcgagggagctaagaagttccctaacaccgagttcgtgaaggtgaagggcctccacttcagccaggaggacgctccagatgaaatgggtaagtacatcaagagcttcgtggagcgcgtgctgaagaacgagcagtaattctaggcgaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 1276 DNA PAT misc_feature 1..1276 note IRES+NanoLuc DNA source 1..1276 mol_type other DNA organism synthetic construct cccggatcctaatacgactcactataggcgcatattaaggtgacgcgtaattcacgcgtcgagcatgcatctagggcggccaattccgcccctctcccccccccccctctccctcccccccccctaacgttactggccgaagccgcttggaataaggccggtgtgcgtttgtctatatgttattttccaccatattgccgtcttttggcaatgtgagggcccggaaacctggccctgtcttcttgacgagcattcctaggggtctttcccctctcgccaaaggaatgcaaggtctgttgaatgtcgtgaaggaagcagttcctctggaagcttcttgaagacaaacaacgtctgtagcgaccctttgcaggcagcggaaccccccacctggcgacaggtgcctctgcggccaaaagccacgtgtataagatacacctgcaaaggcggcacaaccccagtgccacgttgtgagttggatagttgtggaaagagtcaaatggctctcctcaagcgtattcaacaaggggctgaaggatgcccagaaggtaccccattgtatgggatctgatctggggcctcggtgcacatgctttacatgtgtttagtcgaggttaaaaaaacgtctaggccccccgaaccacggggacgtggttttcctttgaaaaacacgatgataagcttgccacaacccgggatcgccaccatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagagtcggggcggccggccgcttaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 4396 DNA PAT misc_feature 1..4396 note pIRES Vector + pNL1.1 TK Vector source 1..4396 mol_type other DNA organism synthetic construct ggcctaactggccggtacctgagtctaaatgagtcttcggacctcgcgggggccgcttaagcggtggttagggtttgtctgacgcggggggagggggaaggaacgaaacactctcattcggaggcggctcggggtttggtcttggtggccacgggcacgcagaagagcgccgcgatcctcttaagcacccccccgccctccgtggaggcgggggtttggtcggcgggtggtaactggcgggccgctgactcgggcgggtcgcgcgccccagagtgtgaccttttcggtctgctcgcagacccccgggcggcgccgccgcggcggcgacgggctcgctgggtcctaggctccatggggaccgtatacgtggacaggctctggagcatccgcacgactgcggtgatattaccggagaccttctgcgggacgagccgggtcacgcggctgacgcggagcgtccgttgggcgacaaacaccaggacggggcacaggtacactatcttgtcacccggaggcgcgagggactgcaggagcttcagggagtggcgcagctgcttcatccccgtggcccgttgctcgcgtttgctggcggtgtccccggaagaaatatatttgcatgtctttagttctatgatgacacaaaccccgcccagcgtcttgtcattggcgaattcgaacacgcagatgcagtcggggcggcgcggtcccaggtccacttcgcatattaaggtgacgcgtaattcacgcgtcgagcatgcatctagggcggccaattccgcccctctcccccccccccctctccctcccccccccctaacgttactggccgaagccgcttggaataaggccggtgtgcgtttgtctatatgttattttccaccatattgccgtcttttggcaatgtgagggcccggaaacctggccctgtcttcttgacgagcattcctaggggtctttcccctctcgccaaaggaatgcaaggtctgttgaatgtcgtgaaggaagcagttcctctggaagcttcttgaagacaaacaacgtctgtagcgaccctttgcaggcagcggaaccccccacctggcgacaggtgcctctgcggccaaaagccacgtgtataagatacacctgcaaaggcggcacaaccccagtgccacgttgtgagttggatagttgtggaaagagtcaaatggctctcctcaagcgtattcaacaaggggctgaaggatgcccagaaggtaccccattgtatgggatctgatctggggcctcggtgcacatgctttacatgtgtttagtcgaggttaaaaaaacgtctaggccccccgaaccacggggacgtggttttcctttgaaaaacacgatgataagcttgccacaacccgggatcgccaccatggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagagtcggggcggccggccgcttcgagcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtaaaatcgataaggatccgtcgaccgatgcccttgagagccttcaacccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgtcttctttatcatgcaactcgtaggacaggtgccggcagcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagcggccgcaaatgctaaaccactgcagtggttaccagtgcttgatcagtgaggcaccgatctcagcgatctgcctatttcgttcgtccatagtggcctgactccccgtcgtgtagatcactacgattcgtgagggcttaccatcaggccccagcgcagcaatgatgccgcgagagccgcgttcaccggcccccgatttgtcagcaatgaaccagccagcagggagggccgagcgaagaagtggtcctgctactttgtccgcctccatccagtctatgagctgctgtcgtgatgctagagtaagaagttcgccagtgagtagtttccgaagagttgtggccattgctactggcatcgtggtatcacgctcgtcgttcggtatggcttcgttcaactctggttcccagcggtcaagccgggtcacatgatcacccatattatgaagaaatgcagtcagctccttagggcctccgatcgttgtcagaagtaagttggccgcggtgttgtcgctcatggtaatggcagcactacacaattctcttaccgtcatgccatccgtaagatgcttttccgtgaccggcgagtactcaaccaagtcgttttgtgagtagtgtatacggcgaccaagctgctcttgcccggcgtctatacgggacaacaccgcgccacatagcagtactttgaaagtgctcatcatcgggaatcgttcttcggggcggaaagactcaaggatcttgccgctattgagatccagttcgatatagcccactcttgcacccagttgatcttcagcatcttttactttcaccagcgtttcggggtgtgcaaaaacaggcaagcaaaatgccgcaaagaagggaatgagtgcgacacgaaaatgttggatgctcatactcgtcctttttcaatattattgaagcatttatcagggttactagtacgtctctcaaggataagtaagtaatattaaggtacgggaggtattggacaggccgcaataaaatatctttattttcattacatctgtgtgttggttttttgtgtgaatcgatagtactaacatacgctctccatcaaaacaaaacgaaacaaaacaaactagcaaaataggctgtccccagtgcaagtgcaggtgccagaacatttctct 48 DNA PAT misc_feature 1..48 note synthetic oligo DNA source 1..48 mol_type other DNA organism synthetic construct cccggatcctaatacgactcactataggcgcatattaaggtgacgcgt 51 DNA PAT misc_feature 1..51 note synthetic oligo DNA 2 source 1..51 mol_type other DNA organism synthetic construct ttttttttttttttttttttttttttttttctagaattacgccagaatgcg 54 RNA PAT misc_feature 1..54 note 54-nt flagment A source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 54 RNA PAT misc_feature 1..54 note 54-nt flagment B misc_feature 44^45 note Two adenosines(A)-(A) are linked by a branching linker. 5'pG islinked to the branched chain that branches off from the mainchain. source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 54 RNA PAT misc_feature 1..54 note 54-nt flagment C misc_feature 44^45 source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 54 RNA PAT misc_feature 1..54 note 54-nt flagment D misc_feature 14^15 note The guanosine(G) and the uridine(U) are linked by a branchinglinker. 5'G is linked to the branched chain that branches offfrom the main chain. source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 54 RNA PAT misc_feature 1..54 note 54-nt flagment E misc_feature 14^15 note The guanosine(G) and the uridine(U) are linked by a branchinglinker. 5'm7G-ppp-G is linked to the branched chain that branchesoff from the main chain.("5'm7G" indicates CAP structure). source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 550 RNA PAT misc_feature 1..550 note 550-nt flagment (IVT synthesized) source 1..550 mol_type other RNA organism synthetic construct ggtcttcacactcgaagatttcgttggggactggcgacagacagccggctacaacctggaccaagtccttgaacagggaggtgtgtccagtttgtttcagaatctcggggtgtccgtaactccgatccaaaggattgtcctgagcggtgaaaatgggctgaagatcgacatccatgtcatcatcccgtatgaaggtctgagcggcgaccaaatgggccagatcgaaaaaatttttaaggtggtgtaccctgtggatgatcatcactttaaggtgatcctgcactatggcacactggtaatcgacggggttacgccgaacatgatcgactatttcggacggccgtatgaaggcatcgccgtgttcgacggcaaaaagatcactgtaacagggaccctgtggaacggcaacaaaattatcgacgagcgcctgatcaaccccgacggctccctgctgttccgagtaaccatcaacggagtgaccggctggcggctgtgcgaacgcattctggcgtaattctagaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 54 RNA PAT misc_feature 1..54 note 54-nt flagment (n=6) misc_feature 44^45 note Two adenosines(A)-(A) are linked by a branching linker.5'm7G-ppp-GmGmAGCGA is linked to the branched chain that branchesoff from the main chain.("5'm7G" indicates CAP structure. "Gm"indicates 2'OMe guanosine) source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 54 RNA PAT misc_feature 1..54 note 54-nt flagment (n=12) misc_feature 44^45 note Two adenosines(A)-(A) are linked by a branching linker.5'm7G-ppp-GmGmAGCGAUAAA is linked to the branched chain thatbranches off from the main chain.("5'm7G" indicates CAPstructure. "Gm" indicates 2'OMe guanosine) source 1..54 mol_type other RNA organism synthetic construct ggcgcatattaaggtgacgcgtgtggcctcgaacaccgagggtaaagccaccat 24 RNA PAT source 1..24 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage ggaacggttacagcaacgctgctg 24 RNA PAT source 1..24 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage gaccgctttgctggaatcgaggaa 24 RNA PAT source 1..24 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage ggataggtgtctttgtcggttaca 24 RNA PAT source 1..24 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage ggatttgcggtgctttgctggaat 51 RNA PAT source 1..51 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT source 1..51 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 2^3 note 2'-methyl guanosine(Gm) ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 51 RNA PAT source 1..51 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 2..4 note LNA misc_feature 19..21 note LNA ggcgtcaccttaatatgcgccaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage ggagcggcgtcaccttaatatgcgcc 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 17..26 note LNA ggagcggcgtcaccttaatatgcgcc 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 22..26 note LNA ggagcggcgtcaccttaatatgcgcc 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 8 note LNA misc_feature 10 note LNA misc_feature 12 note LNA misc_feature 14 note LNA misc_feature 16 note LNA misc_feature 18 note LNA misc_feature 20 note LNA misc_feature 22 note LNA misc_feature 24 note LNA misc_feature 26 note LNA ggagcggcgtcaccttaatatgcgcc 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 16 note LNA misc_feature 18 note LNA misc_feature 20 note LNA misc_feature 22 note LNA misc_feature 24 note LNA misc_feature 26 note LNA ggagcggcgtcaccttaatatgcgcc 26 RNA PAT source 1..26 mol_type mRNA organism artificial sequence misc_feature 1 note 5' m7g misc_feature 1^2 note triphosphate linkage misc_feature 9 note LNA misc_feature 12 note LNA misc_feature 15 note LNA misc_feature 18 note LNA misc_feature 21 note LNA misc_feature 24 note LNA ggagcggcgtcaccttaatatgcgcc