pZAG1
Designed and
constructed by Yuji Hiwatashi
Construction date: 12 Feb., 2003 Correction date: 2 July, 2012 Comments and Reference: mGFP (start codon: ATG ->TTG) expression vector under control of rice actin promoter, zeocin resistant, pBluescriptKSII. BS213 fragments are used as a platform for integration. 5’UTR+Start codon represents a cDNA fragment encoding a kinesin-like protein. This fragment covers from 5’UTR to the 30th amino acid. You have to replace this fragment with your gene to express the GFP fusion protein.
Pact-rev1:
5’- GGAGGAGCTCGTCGTCCGGCA-3’
The following primer is located upstream of BS213-5’
region. You can check the targeting of
pZAG1 by PCR with 213F2 and Pact-rev1 primers (amplification of ca. 1.3-kb
fragment).
213F2:
5’- AAATCTCATAGGCGTTGCTTG-3’
Sequence:
>pZAG1
TCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTCCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGGTACCATCAGACGTAAATATAAAGTTGTAAGGAAAATTAGTTCTATAAAATGCCTCTATATTAACATTCAATTGTAAAGGTGCTGCGTATTCATGGATTAGTATATAAATTATTATTTTTGTAAAATGTGTGTGTGTGTTGTTAATGCTTTATGTAAAAAGAACTGTGTGCTTCAAAATACTATAGATCCCTTTTAGACAGATTATCTTTATGTAAAAAGAACTGTGTGCTTCAAAATACTATAGATCCCTTTTAGACAGAATATCTATAATATGTAGAAGTTAAATTCGGTTTTAATTCTGTACCATGTATTGTGAACATCTAGATTTTGTGATACAGCTTCATGAAAAACTTCAAAACTTCAAAACATTTTACTTCTATAAAGTTATCAATTTGTTTGACAAACAACCTTATATGGTAATGACAAGAGTCTCACATTAATTCAGCATCTCAAAAATAAATATAAATACTCTTTTATACTCAAAATTGTATTTAATACAAAACATATGCATGTATCAATCCAATGACTTGGGCCAATAATGTAAGTCACTCATTATAGTATTAGTATTTATTATTCCTTTTTGTTTCTCAGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACTCATCTATGTGATTCGTTCTCAACACTTACACTTCAAAAATATTTATTTACTCATTCATCAATACATCCAAGATAAAATATATATATATAGATATATATATATATATATATATATATTTATCTTTTTCTAACCTCTTCTACAGAGTTTTGTTCAGTGTTGTAAACCCAAACTTGGTGTCGAGGGCCCGAGGTCATTCATATGCTTGAGAAGAGAGTCGGGATAGTCCAAAATAAAACAAAGGTAAGATTACCTGGTCAAAAGTGAAAACATCAGTTAAAAGGTGGTATAAAGTAAAATATCGGTAATAAAAGGTGGCCCAAAGTGAAATTTACTCTTTTCTACTATTATAAAAATTGAGGATGTTTTTGTCGGTACTTTGATACGTCATTTTTGTATGAATTGGTTTTTAAGTTTATTCGCTTTTGGAAATGCATATCTGTATTTGAGTCGGGTTTTAAGTTCGTTTGCTTTTGTAAATACAGAGGGATTTGTATAAGAAATATCTTTAAAAAAACCCATATGCTAATTTGACATAATTTTTGAGAAAAATATATATTCAGGCGAATTCTCACAATGAACAATAATAAGATTAAAATAAGCTTTCCCCCGTTGCAGCGCATGGGTATTTTTTCTAGTAAAAATAAAAGATAAACTTAGACTCAAAACATTTACAAAAACAACCCCTAAAGTTCCTAAAGCCCAAAGTGCTATCCACGATCCATAGCAAGCCCAGCCCAACCCAACCCAACCCAACCCACCCCAGTCCAGCCAACTGGACAATAGTCTCCACACCCCCCCACTATCACCGTGAGTTGTCCGCACGCACCGCACGTCTCGCAGCCAAAAAAAAAAAAGAAAGAAAAAAAAGAAAAAGAAAAAACAGCAGGTGGGTCCGGGTCGTGGGGGCCGGAAACGCGAGGAGGATCGCGAGCCAGCGACGAGGCCGGCCCTCCCTCCGCTTCCAAAGAAACGCCCCCCATCGCCACTATATACATACCCCCCCCTCTCCTCCCATCCCCCCAACCCTACCACCACCACCACCACCACCTCCACCTCCTCCCCCCTCGCTGCCGGACGACGAGCTCCTCCCCCCTCCCCCTCCGCCGCCGCCGCGCCGGTAACCACCCCGCCCCTCTCCTCTTTCTTTCTCCGTTTTTTTTTTCCGTCTCGGTCTCGATCTTTGGCCTTGGTAGTTTGGGTGGGCGAGAGGCGGCTTCGTGCGCGCCCAGATCGGTGCGCGGGAGGGGCGGGATCTCGCGGCTGGGGCTCTCGCCGGCGTGGATCCGGCCCGGATCTCGCGGGGAATGGGGCTCTCGGATGTAGATCTGCGATCCGCCGTTGTTGGGGGAGATGATGGGGGGTTTAAAATTTCCGCCATGCTAAACAAGATCAGGAAGAGGGGAAAAGGGCACTATGGTTTATATTTTTATATATTTCTGCTGCTTCGTCAGGCTTAGATGTGCTAGATCTTTCTTTCTTCTTTTTGTGGGTAGAATTTGAATCCCTCAGCATTGTTCATCGGTAGTTTTTCTTTTCATGATTTGTGACAAATGCAGCCTCGTGCGGAGCTTTTTTGTAGGTAGAAGATATCGGGGATCCTCTAGAGTCGACGAAGCACCTCGCTCAATGTCTCTAAAACGTCAAGCCCCTCAGTCCTGTGGAATGAAACTCTTCTAGCGACCCCAGTGTAGTGTGACAGAGGGCTGGTGAGGACGAGGATTCCAAGAATGTCGTCTTGCTTATGCAGGTAAGGAGCTTATCCTCCAGACATCGACTAGAGGATCCATGCACCTATCATCTGGAATTTGAAATCCACTTTCTTACAGTGTCCTCTACATTGCGGCCTTCACGGTGACCGCTGGAGTGTGCCCACATACTCCCTTAAAACAGAGCCGTCGGGTGCGATGACAACGATGATGTCCCCGAAGCCGAGGGAGAAGAGGCCGTCTGGCAATGTAGGGGGTAATACACCCACGGGTAAGGCCAGGTCTATCGATAAGCTTTTGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCTTCACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCACGGCATGGACGAGCTGTACAAGTAAAGCGGCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCCGATGATAAGCTGTCAAACATGAGAATTCCCTTTCAGAAAGAATGCTAACCCACAGATGGTTAGAGAGGCTTACGCAGCAGGTCTCATCAAGACGATCTACCCGAGCAATAATCTCCAGGAAATCAAATACCTTCCCAAGAAGGTTAAAGATGCAGTCAAAAGATTCAGGACTAACTGCATCAAGAACACAGAGAAAGATATATTTCTCAAGATCAGAAGTACTATTCCAGTATGGACGATTCAAGGCTTGCTTCACAAACCAAGGCAAGTAATAGAGATTGGAGTCTCTAAAAAGGTAGTTCCCACTGAATCAAAGGCCATGGAGTCAAAGATTCAAATAGAGGACCTAACAGAACTCGCCGTAAAGACTGGCGAACAGTTCATACAGAGTCTCTTACGACTCAATGACAAGAAGAAAATCTTCGTCAACATGGTGGAGCACGACACACTTGTCTACTCCAAAAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCAATTGAGACTTTTCAACAAAGGGTAATATCCGGAAACCTCCTCGGATTCCATTGCCCAGCTATCTGTCACTTTATTGTGAAGATAGTGGAAAAGGAAGGTGGCTCCTACAAATGCCATCATTGCGATAAAGGAAAGGCCATCGTTGAAGATGCCTCTGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGAAAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATATCTCCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACCCTTCCTCTATATAAGGAAGTTCATTTCATTTGGAGAGGGGATCCATGGCCAAGTTGACCAGTGCCGTTCCGGTGCTCACCGCGCGCGACGTCGCCGGAGCGGTCGAGTTCTGGACCGACCGGCTCGGGTTCTCCCGGGACTTCGTGGAGGACGACTTCGCCGGTGTGGTCCGGGACGACGTGACCCTGTTCATCAGCGCGGTCCAGGACCAGGTGGTGCCGGACAACACCCTGGCCTGGGTGTTGGTGCGCGGCCTGGACGAGCTGTACGCCGAGTGGTCGGAGGTCGTGTCCACGAACTTCCGGGACGCCTCCGGGCCGGCCATGACCGAGATCGGCGAGCAGCCGTGGGGGCGGGAGTTCGCCCTGCGCGACCCGGCCGGCAACTGCGTGCACTTCGTGGCCGAGGAGCAGGACTGAGCGGGACTCTGGGGTTCGGACGGTACGCTGAAATCACCAGTCTCTCTCTACAAATCTATCTCTCTCTATTTTCTCCATAAATAATGTGTGAGTAGTTTCCCGATAAGGGAAATTAGGGTTCTTATAGGGTTTCGCTCATGTGTTGAGCATATAAGAAACCCTTAGTATGTATTTGTATTTGTAAAATACTTCTATCAATAAAATTTCTAATTCCTAAAACCAAAATCCAGTACTAAAATCCAGATCTCCTAAAGTCCCTATAGATCTTTGTCGTGAATATAAACCAGACACGAGACGACTAAACCTGGAGCCCAGACGCCGTTCGAAGCTAGAAGTACCGCTTAGGCAGGAGGCCGTTAGGGAAAAGATGCTAAGGCAGGGTTGGTTACGTTGACTCCCCCGTAGGTTTGGTTTAAATATGATGAAGTGGACGGAAGGAAGGAGGAAGACAAGGAAGGATAAGGTTGCAGGCCCTGTGCAAGGTAAGAAGATGGAAATTTGATAGAGGTACGCTACTATACTTATACTATACGCTAAGGGAATGCTTGTATTTATACCCTATACCCCCTAATAACCCCTTATCAATTTAAGAAATAATCCGCATAAGCCCCCGCTTAAAAATTGGTATCAGAGCCATGAATAGGTCTATGACCAAAACTCAAGAGGATAAAACCTCACCAAAATACGAAAGAGTTCTTAACTCTAAAGATAAAAGATCTTTCAAGATCAAAACTAGTTCCCTCACACCGGTGACGGGGATCCACTAGTTCTAGAGCGGCCGCAACTTAGAAGTAGCGTGTTTCTCTACATCACGCTTTTATCCCTTTGTAAATTTTAGAGCGTAGGTTGGCATGGGGGGGCTATTGGCTGTGAAAGGGACAGAGATATCTGCTGCACCATGTTCTGTACATAAAGCTGCCGCTCTGTTACGCCGTTTCTTGTGAACCGAGGCAGCTTCTGAGGTTGGCTTTGCTTAAAGTTGCGCTGATGCTGTTTCCCAATGTTTAGAGGAAATCCTGGTGACGAGAACGAAGAACTCGGGAATTACTAGGGAGGATTTGAGGTTTGTATCTGCAGGGCGTGGTTCAAAATTCTTTCATGTCGAACAGGTTGACAAAGTTCCAGAAATTATGAAATCTCAAAGAGGTAAAGGGTCGAAACGTTTGAAGATCTCGAAAGAAAAAGGAGGAGAAAGGTTGAACATCATTTCATTTTAAGGCATTTCGAGGACATAAAATTGGATGGTGCAGAGGGGAGAAAGCAGGCCAATGGGAAGCTAGGTGTCGATGAAAAGCCTGTCGTTACCGAATTGCTTTCGAGTACACCTAGGCAATGAGATCCAGAAGATAATTACAGAGAATTTCAAAAGAATGAAAAGATGAAGAAAAAGAAGAAGAGTTTGAATGGAGAGGCAATGGATGTGGATGTAGGAGGGTTGTACTCGGACATTTTGATCGAGGCCGATGTGGATTGGTACCCTGATATTACGAAAAACATTGTAGAGCATCTGACATAATGTCTTCTAAAGTACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAG