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== Fusion Protein == We need to fusion protein blah blah blah {{:Team:NYMU-Taipei/NStyle}} === GFP === We based splitting GFP ({{:Team:NYMU-Taipei/BBa|E0040}}) based on the split point at 157&158aa used by iGEM07_Davidson_Missouri's {{:Team:NYMU-Taipei/BBa|I715019}} and {{:Team:NYMU-Taipei/BBa|I715020}}. The A-part split is the same as {{:Team:NYMU-Taipei/BBa|I715019}}, but the B-part has one base difference from {{:Team:NYMU-Taipei/BBa|I715020}}. {{:Team:NYMU-Taipei/Seq|GFP ({{:Team:NYMU-Taipei/BBa|E0040}})|720| atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggaga gggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactt tcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaa agaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaagg tattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaaga atggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgat ggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttga gtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa }} {{:Team:NYMU-Taipei/Seq|GFP_A ({{:Team:NYMU-Taipei/BBa|I175019}})|471| {{:Team:NYMU-Taipei/N|FP|atgcgtaaaggagaagaacttttc}}actggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggaga gggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactt tcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaa agaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaagg tattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaat{{:Team:NYMU-Taipei/N|RP|gtatacatcatggcagacaaacaa}} }} FP (including start codon): atgcgtaaaggagaagaacttttc (55c,24bp,38gc) RP: ttgtttgtctgccatgatgtatac (55c,24bp,38gc) {{:Team:NYMU-Taipei/Seq|GFP_B|249| {{:Team:NYMU-Taipei/N|FP|aagaatggaatcaaagttaacttcaaaa}}ttagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattg gcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtcctt cttgagtttgtaacagctgctgggattaca{{:Team:NYMU-Taipei/N|RP|catggcatggatgaactatacaaa}}taataa }} FP: aagaatggaatcaaagttaacttcaaaa (55c,28bp,25gc) RP (excluding stop codon): tttgtatagttcatccatgccatg (55c,24bp,38gc) === RFP === We based splitting RFP ({{:Team:NYMU-Taipei/BBa|E1010}}) based on the split point at 154&155aa used by iGEM07_Davidson_Missouri's {{:Team:NYMU-Taipei/BBa|I715022}} and {{:Team:NYMU-Taipei/BBa|I715023}}. The A-part split is the same as {{:Team:NYMU-Taipei/BBa|I715022}}, but the B-part has one base difference from {{:Team:NYMU-Taipei/BBa|I715023}}. {{:Team:NYMU-Taipei/Seq|RFP ({{:Team:NYMU-Taipei/BBa|E1010}})|681| atggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtg aaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagta cggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaa gacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtc cggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggtgctctgaaaggtgaaatcaaaatgcgtctgaaact gaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggac atcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataa }} {{:Team:NYMU-Taipei/Seq|RFP_A ({{:Team:NYMU-Taipei/BBa|I715022}})|462| {{:Team:NYMU-Taipei/N|FP|atggcttcctccgaagac}}gttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtg aaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagta cggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaa gacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtc cggttatgcagaaaaaaaccatgggttgggaagcttccaccg{{:Team:NYMU-Taipei/N|RP|aacgtatgtacccggaagac}} }} FP (including start codon): atggcttcctccgaagac (55C,18bp,56%) RP: gtcttccgggtacatacgtt (55C,20bp,50%) {{:Team:NYMU-Taipei/Seq|RFP_B|219| {{:Team:NYMU-Taipei/N|FP|ggtgctctgaaaggtgaaatc}}aaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttc agctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcg{{:Team:NYMU-Taipei/N|RP|tca ctccaccggtgct}}taataa }} FP: ggtgctctgaaaggtgaaatc (55C,21bp,48%) RP (excluding stop codon): agcaccggtggagtga (56C,16bp,63%) === eIF4A === We split eIF4A ({{:Team:NYMU-Taipei/BBa|K411100}}) at 215&216aa based on [http://igem.ym.edu.tw/pv2010/images/b/be/File:NYMU_Nmeth1023-S5.pdf 2.protocol file from Natalia E. Broude]. But first we needed to mutate the two PstI cutting sites. We took the protein coding region from the [http://www.ncbi.nlm.nih.gov/nuccore/NM_144958 eIF4A mRNA transcript sequence from Mouse (from NCBI)] and found it had 2 PstI cutting sites: {{:Team:NYMU-Taipei/Seq|eIF4A_original([http://partsregistry.org/Part:BBa_K411100 BBa_K411100])|1173| atggagccggaaggcgtcatcgagagtaactggaacgagattgtggatagctttgatgacatgaatctctcagagtccctcctccgtggtatttatgcct atggttttgagaagccctctgccatccagcagcgagctattcttccttgtatcaagggttatgatgtgattgctcaagcccagtctgggactgggaaaac agctacatttgccatatcaatt{{:Team:NYMU-Taipei/N|cut|ctgcag}}cagattgaattagatctaaaggccactcaggctttggttctggcacccacacgtgaattggctcagcagata caaaaggtggttatggcattaggagactacatgggtgcctcttgtcatgcctgcattgggggcaccaatgtgcgtgctgaggtgcagaag{{:Team:NYMU-Taipei/N|cut|ctgcag}}atgg aagctccccatatcatcgtgggtacccctggccgggtgtttgacatgcttaaccggagatacctgtcccccaaatacatcaagatgttcgtactggatga agcagatgaaatgttaagccgagggttcaaggatcagatctatgacatattccagaagctcaacagcaacacacaggtagttttgttgtctgctacaatg ccttctgatgtccttgaggtgaccaagaaatttatgagagaccctattcggattcttgtcaagaaggaagaattgaccctggagggtatccgccaattct acatcaatgtggaacgagaggagtggaagcttgacacattgtgtgacttgtatgagacgctgaccatcacccaggcagtcatctttatcaacaccagaag gaaggtggactggctcaccgagaagatgcatgcccgagatttcactgtttctgccatgcacggagatatggaccaaaaggaacgagatgtgatcatgagg gagttccggtctggctctagcagagtattaattaccactgacctgttggccagaggcattgatgtgcagcaggtctccttagtcatcaactatgaccttc ccaccaacagggaaaactacatccacagaatcggtcgaggtggtcggtttggtcgtaagggtgtggctattaacatggtgaccgaagaagacaagaggac tcttcgagacattgagactttctacaacacctccattgaagagatgcccctcaacgttgctgacctcatttga }} We need to mutate out the two PstI cutting sites. The template of eIF4A on a [http://genome-www.stanford.edu/vectordb/vector_descrip/COMPLETE/PGEX4T1.SEQ.html pGEX-4TI vector] was kindly provided by <font color="red">C.Proud</font>. We used these primers to mutate the cutting site: <font color="blue">2 of pstI cutting site</font> mut1(24bp,55C,42%) FP: ccatatcaatt<font color="red">ctccag</font>cagattga RP: caatctg<font color="red">ctggag</font>aattgatatggc mut2(18bp,55C,56%) FP: gcagaag<font color="red">ctccag</font>atggaa RP: tccat<font color="red">ctggag</font>cttctgca The new sequence after mutation: {{:Team:NYMU-Taipei/Seq|eIF4A ({{:Team:NYMU-Taipei/BBa|K411100}})|1173| atggagccggaaggcgtcatcgagagtaactggaacgagattgtggatagctttgatgacatgaatctctcagagtccctcctccgtggtatttatgcct atggttttgagaagccctctgccatccagcagcgagctattcttccttgtatcaagggttatgatgtgattgctcaagcccagtctgggactgggaaaac agctacatttg{{:Team:NYMU-Taipei/N|OP|ccatatcaatt{{:Team:NYMU-Taipei/N|cutmut|ctccag}}cagattga}}attagatctaaaggccactcaggctttggttctggcacccacacgtgaattggctcagcagata caaaaggtggttatggcattaggagactacatgggtgcctcttgtcatgcctgcattgggggcaccaatgtgcgtgctgaggt{{:Team:NYMU-Taipei/N|OP|gcagaag{{:Team:NYMU-Taipei/N|cutmut|ctccag}}atgg aa}}gctccccatatcatcgtgggtacccctggccgggtgtttgacatgcttaaccggagatacctgtcccccaaatacatcaagatgttcgtactggatga agcagatgaaatgttaagccgagggttcaaggatcagatctatgacatattccagaagctcaacagcaacacacaggtagttttgttgtctgctacaatg ccttctgatgtccttgaggtgaccaagaaatttatgagagaccctattcggattcttgtcaagaaggaagaattgaccctggagggtatccgccaattct acatcaatgtggaacgagaggagtggaagcttgacacattgtgtgacttgtatgagacgctgaccatcacccaggcagtcatctttatcaacaccagaag gaaggtggactggctcaccgagaagatgcatgcccgagatttcactgtttctgccatgcacggagatatggaccaaaaggaacgagatgtgatcatgagg gagttccggtctggctctagcagagtattaattaccactgacctgttggccagaggcattgatgtgcagcaggtctccttagtcatcaactatgaccttc ccaccaacagggaaaactacatccacagaatcggtcgaggtggtcggtttggtcgtaagggtgtggctattaacatggtgaccgaagaagacaagaggac tcttcgagacattgagactttctacaacacctccattgaagagatgcccctcaacgttgctgacctcatttga }} {{:Team:NYMU-Taipei/Seq|eIF4A_A|645| {{:Team:NYMU-Taipei/N|FP|atggagccggaaggcgtcatcga}}gagtaactggaacgagattgtggatagctttgatgacatgaatctctcagagtccctcctccgtggtatttatgcct atggttttgagaagccctctgccatccagcagcgagctattcttccttgtatcaagggttatgatgtgattgctcaagcccagtctgggactgggaaaac agctacatttgccatatcaattctccagcagattgaattagatctaaaggccactcaggctttggttctggcacccacacgtgaattggctcagcagata caaaaggtggttatggcattaggagactacatgggtgcctcttgtcatgcctgcattgggggcaccaatgtgcgtgctgaggtgcagaagctccagatgg aagctccccatatcatcgtgggtacccctggccgggtgtttgacatgcttaaccggagatacctgtcccccaaatacatcaagatgttcgtactggatga agcagatgaaatgttaagccgagggttcaaggatcagatctatgacatattccagaagctcaacagcaacacacaggtagttttgttgtctgctacaatg ccttctgatgtccttgaggtga{{:Team:NYMU-Taipei/N|RP|ccaagaaatttatgagagaccct}} }} FP: atggagccggaaggcgtcatcga (66c,24bp,61gc) [design error -- was suppose to design it to be 55c] RP: agggtctctcataaatttcttgg (54C,23bp,39%) {{:Team:NYMU-Taipei/Seq|eIF4A_B|528| {{:Team:NYMU-Taipei/N|FP|attcggattcttgtcaagaagg}}aagaattgaccctggagggtatccgccaattct acatcaatgtggaacgagaggagtggaagcttgacacattgtgtgacttgtatgagacgctgaccatcacccaggcagtcatctttatcaacaccagaag gaaggtggactggctcaccgagaagatgcatgcccgagatttcactgtttctgccatgcacggagatatggaccaaaaggaacgagatgtgatcatgagg gagttccggtctggctctagcagagtattaattaccactgacctgttggccagaggcattgatgtgcagcaggtctccttagtcatcaactatgaccttc ccaccaacagggaaaactacatccacagaatcggtcgaggtggtcggtttggtcgtaagggtgtggctattaacatggtgaccgaagaagacaagaggac tcttcgagacattgagactttctacaacacctccattgaagagatgccc{{:Team:NYMU-Taipei/N|RP|ctcaacgttgctgacctcatttga}} }} FP: attcggattcttgtcaagaagg (54C,22bp,41%) RP (including stop codon): tcaaatgaggtcagcaacgttgag (59c,24bp,46gc) [design error -- was suppose to design it to be 55c] === Linker === We took the linker from 2.protocol file from Natalia E. Broude:[http://igem.ym.edu.tw/pv2010/images/b/be/File:NYMU_Nmeth1023-S5.pdf ] {{:Team:NYMU-Taipei/Seq|linker GSSGSSGSGS|30| ggcagcagcggcagcagcggcagcggcagc }} We decided to use primers to create the linker: cggcagcagcggcagcggcagc ggcagcagcggcagcagcggcagcggcagc ggcagcagcggcagcagcggcagc overlapping region: cggcagcagcggcagc (63c,16bp,81gc) FP: cggcagcagcggcagcggcagc (22bp) RP: gctgccgctgctgccgctgctgcc (24bp) === Fusioning it together === ==== GFP fusion part ==== (note the extra taa stop codon at the end) {{:Team:NYMU-Taipei/Seq|GFP-linker-eIF4A_A ({{:Team:NYMU-Taipei/BBa|K411101}})|1149| {{:Team:NYMU-Taipei/N|FP|atgcgtaaaggagaagaacttttc}}actggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtg aaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatgg tgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttc aaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatg gaaacattcttggacacaaattggaatacaactataactcacacaat{{:Team:NYMU-Taipei/N|RP|gtatacatcatggcagacaaacaa}}{{:Team:NYMU-Taipei/N|S|{{:Team:NYMU-Taipei/N|RP|ggcagcag}}{{:Team:NYMU-Taipei/N|OP|cggcagcagcggcagc}}{{:Team:NYMU-Taipei/N|FP|ggcag c}}}}{{:Team:NYMU-Taipei/N|FP|atggagccggaaggcgtcatcga}}gagtaactggaacgagattgtggatagctttgatgacatgaatctctcagagtccctcctccgtggtatttatgcc tatggttttgagaagccctctgccatccagcagcgagctattcttccttgtatcaagggttatgatgtgattgctcaagcccagtctgggactgggaaaa cagctacatttgccatatcaattctccagcagattgaattagatctaaaggccactcaggctttggttctggcacccacacgtgaattggctcagcagat acaaaaggtggttatggcattaggagactacatgggtgcctcttgtcatgcctgcattgggggcaccaatgtgcgtgctgaggtgcagaagctccagatg gaagctccccatatcatcgtgggtacccctggccgggtgtttgacatgcttaaccggagatacctgtcccccaaatacatcaagatgttcgtactggatg aagcagatgaaatgttaagccgagggttcaaggatcagatctatgacatattccagaagctcaacagcaacacacaggtagttttgttgtctgctacaat gccttctgatgtccttgaggtga{{:Team:NYMU-Taipei/N|RP|ccaagaaatttatgagagacccttga}} }} GFP_split_A_FP: gaattcgcggccgcttctagag atgcgtaaaggagaagaacttttc (46bp) OK GFP_split_A_RP: gctgccgctgctgccgctgctgcc ttgtttgtctgccatgatgtatac (48bp) RE DING eIF4A_split_A_FP: cggcagcagcggcagcggcagc atggagccggaaggcgtcatcga (45bp) OK eIF4A_split_A_RP: ctgcagcggccgctactagta tca agggtctctcataaatttcttgg (47bp) OK (note that we added as start codon and removed the stop codon of GFP_B) {{:Team:NYMU-Taipei/Seq|GFP-linker-eIF4A_B ({{:Team:NYMU-Taipei/BBa|K411102}})|804| {{:Team:NYMU-Taipei/N|FP|atgaagaatggaatcaaagttaacttcaaaa}}ttagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcg atggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtt tgtaacagctgctgggattaca{{:Team:NYMU-Taipei/N|RP|catggcatggatgaactatacaaa}}{{:Team:NYMU-Taipei/N|S|{{:Team:NYMU-Taipei/N|RP|ggcagcag}}{{:Team:NYMU-Taipei/N|OP|cggcagcagcggcagc}}{{:Team:NYMU-Taipei/N|FP|ggcagc}}}}{{:Team:NYMU-Taipei/N|FP|attcggattcttgtcaagaagg}}aa gaattgaccctggagggtatccgccaattctacatcaatgtggaacgagaggagtggaagcttgacacattgtgtgacttgtatgagacgctgaccatca cccaggcagtcatctttatcaacaccagaaggaaggtggactggctcaccgagaagatgcatgcccgagatttcactgtttctgccatgcacggagatat ggaccaaaaggaacgagatgtgatcatgagggagttccggtctggctctagcagagtattaattaccactgacctgttggccagaggcattgatgtgcag caggtctccttagtcatcaactatgaccttcccaccaacagggaaaactacatccacagaatcggtcgaggtggtcggtttggtcgtaagggtgtggcta ttaacatggtgaccgaagaagacaagaggactcttcgagacattgagactttctacaacacctccattgaagagatgccc{{:Team:NYMU-Taipei/N|RP|ctcaacgttgctgacctcat ttga}} }} GFP_split_B_FP: gaattcgcggccgcttctagag atg aagaatggaatcaaagttaacttcaaaa (53bp) GFP_split_B_RP: gctgccgctgctgccgctgctgcc tttgtatagttcatccatgccatg (48bp) eIF4A_split_B_FP: cggcagcagcggcagcggcagc attcggattcttgtcaagaagg (44bp) eIF4A_split_B_RP: ctgcagcggccgctactagta tcaaatgaggtcagcaacgttgag (45bp) ==== RFP fusion part ==== (note the extra taa stop codon at the end) {{:Team:NYMU-Taipei/Seq|RFP-linker-eIF4A_A ({{:Team:NYMU-Taipei/BBa|K411103}})|1140| {{:Team:NYMU-Taipei/N|FP|atggcttcctccgaagac}}gttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtg aaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagta cggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaa gacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtc cggttatgcagaaaaaaaccatgggttgggaagcttccaccg{{:Team:NYMU-Taipei/N|RP|aacgtatgtacccggaagac}}{{:Team:NYMU-Taipei/N|S|{{:Team:NYMU-Taipei/N|RP|ggcagcag}}{{:Team:NYMU-Taipei/N|OP|cggcagcagcggcagc}}{{:Team:NYMU-Taipei/N|FP|ggcagc}}}}{{:Team:NYMU-Taipei/N|FP|atggagcc ggaaggcgtcatcga}}gagtaactggaacgagattgtggatagctttgatgacatgaatctctcagagtccctcctccgtggtatttatgcctatggtttt gagaagccctctgccatccagcagcgagctattcttccttgtatcaagggttatgatgtgattgctcaagcccagtctgggactgggaaaacagctacat ttgccatatcaattctccagcagattgaattagatctaaaggccactcaggctttggttctggcacccacacgtgaattggctcagcagatacaaaaggt ggttatggcattaggagactacatgggtgcctcttgtcatgcctgcattgggggcaccaatgtgcgtgctgaggtgcagaagctccagatggaagctccc catatcatcgtgggtacccctggccgggtgtttgacatgcttaaccggagatacctgtcccccaaatacatcaagatgttcgtactggatgaagcagatg aaatgttaagccgagggttcaaggatcagatctatgacatattccagaagctcaacagcaacacacaggtagttttgttgtctgctacaatgccttctga tgtccttgaggtga{{:Team:NYMU-Taipei/N|RP|ccaagaaatttatgagagacccttga}} }} RFP_split_A_FP: gaattcgcggccgcttctag atggcttcctccgaagac (38bp) RFP_split_A_RP: gctgccgctgctgccgctgctgcc gtcttccgggtacatacgtt (44bp) eIF4A_split_A_FP and RP are the same as the GFP part above. (note that we added as start codon and removed the stop codon of RFP_B) {{:Team:NYMU-Taipei/Seq|RFP-linker-eIF4A_B ({{:Team:NYMU-Taipei/BBa|K411104}})|774| {{:Team:NYMU-Taipei/N|FP|atgggtgctctgaaaggtgaaatc}}aaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccgg ttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcg {{:Team:NYMU-Taipei/N|RP|tcactccaccggtgct}}{{:Team:NYMU-Taipei/N|S|{{:Team:NYMU-Taipei/N|RP|ggcagcag}}{{:Team:NYMU-Taipei/N|OP|cggcagcagcggcagc}}{{:Team:NYMU-Taipei/N|FP|ggcagc}}}}{{:Team:NYMU-Taipei/N|FP|attcggattcttgtcaagaagg}}aagaattgaccctggagggtatccgccaattc tacatcaatgtggaacgagaggagtggaagcttgacacattgtgtgacttgtatgagacgctgaccatcacccaggcagtcatctttatcaacaccagaa ggaaggtggactggctcaccgagaagatgcatgcccgagatttcactgtttctgccatgcacggagatatggaccaaaaggaacgagatgtgatcatgag ggagttccggtctggctctagcagagtattaattaccactgacctgttggccagaggcattgatgtgcagcaggtctccttagtcatcaactatgacctt cccaccaacagggaaaactacatccacagaatcggtcgaggtggtcggtttggtcgtaagggtgtggctattaacatggtgaccgaagaagacaagagga ctcttcgagacattgagactttctacaacacctccattgaagagatgccc{{:Team:NYMU-Taipei/N|RP|ctcaacgttgctgacctcatttga}} }} RFP_split_B_FP: gaattcgcggccgcttctag atg ggtgctctgaaaggtgaaatc (44bp) RFP_split_B_RP: gctgccgctgctgccgctgctgcc agcaccggtggagtga (40bp) eIF4A_split_B_FP and RP are the same as the GFP part above. == Aptamer == * Aptamer in front of non-coding sequence: ** [[Image:NYMU Aptamer Structure with bb scars non-coding predicted by RNAfold.png|frame|none|RNAfold prediction of Aptamer Structure with Biobrick scars in front of a non-coding region. The scars bond together, but the overall structure does not change.]] * Check to see if the addition of the biobrick scars affect the secondary structure in front of coding sequence: ** [[Image:NYMU Aptamer Structure with bb scars coding predicted by RNAfold.png|frame|none|RNAfold prediction of Aptamer Structure with Biobrick scars in front of a coding region. The scars bond together, but the overall structure does not change.]] * Structure interacting with the split proteins: [[Image:NYMU Aptamer Structure interacting with split protein from Paper.png|frame|none| Aptamer Structure interacting with split protein [6]]] {{:Team:NYMU-Taipei/Footer}}

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