Difference between revisions of "Part:BBa K4158004:Design"

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1. Prepare pUC19 cloning vector and this part.
 
1. Prepare pUC19 cloning vector and this part.
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2. Amplitude pUC19 cloning vector with PCR using these Primers Fw:5'-TCTAGAGTCGACCTGCAGG-3' Rv:5'-CCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAAC-3. Amplitude this part with PCR using These Primers Fw:
+
 
 +
2. Amplitude pUC19 cloning vector with PCR using these Primers below.
 +
 
 +
  Fw:5'-CTCTAGAGTCGACCTGCAGG-3'  
 +
 
 +
  Rv:5'-TTAGAATTCAGCCGAGATCTTTTCTAGAGCCGCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAAC-3'
 +
 
 +
3. Amplitude this part with PCR using These Primers  
 +
 
 +
  Fw:5'-GGCTGAATTCTAAAGATCTCATGCGAGT-3'
 +
 
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  Rv:5'-TCGACTCTAGAGGATCTCGATCCGGATA-3'
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4. Restriction and insertion cloning with these PCR productions and restriction enzymes EcoRI and XbaI
  
 
===Source===
 
===Source===

Latest revision as of 07:06, 11 September 2022


PatzR-sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4
    Illegal XhoI site found at 115
    Illegal XhoI site found at 587
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 977
    Illegal BsaI site found at 1155


Design Notes

If you want to get same plasmid that we experimented with, construct the plasmid along this way below.

1. Prepare pUC19 cloning vector and this part.

2. Amplitude pUC19 cloning vector with PCR using these Primers below. 

  Fw:5'-CTCTAGAGTCGACCTGCAGG-3' 

  Rv:5'-TTAGAATTCAGCCGAGATCTTTTCTAGAGCCGCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAAC-3'

3. Amplitude this part with PCR using These Primers 

  Fw:5'-GGCTGAATTCTAAAGATCTCATGCGAGT-3'

  Rv:5'-TCGACTCTAGAGGATCTCGATCCGGATA-3'

4. Restriction and insertion cloning with these PCR productions and restriction enzymes EcoRI and XbaI

Source

TAAAGATCTCATGCGAGTCAAAGCAAGATCGGTGCCGGATCGGCACCAGTTAGGTCGGAAAAAGGCGGCAGTCAAGTGCGCAGGGCGGCGTTATAATTGAACGAAACGTCGACTCTCGAGTGAGATTGTTGACGGTACCGTATTTTGGATCTAGGAGGAAGGATCTATGAGCAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCCGTGGAGAGGGTGAAGGTGATGCTACAAACGGAAAACTCACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCGTGGCCAACACTTGTCACTACTCTGACCTATGGTGTTCAATGCTTTTCCCGTTATCCGGATCACATGAAACGGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAACGCACTATATCTTTCAAAGATGACGGGACCTACAAGACGCGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATCGTATCGAGTTAAAGGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAACTCGAGTACAACTTTAACTCACACAATGTATACATCACGACAGACAAACAAAAGAATGGAATCAAAGCTAACTTCAAAATTCGCCACAACGTTGAAGATGGTTCCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCGACACAATCTGTCCTTTCGAAAGATCCCAACGAAAAGCGTGACCACATGGTCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAGCTCTACAAATAAGGATCTGAAGCTTGGGCCCGAACAAAAACTCATCTCAGAAGAGGATCTGAATAGCGCCGTCGACCATCATCATCATCATCATTGAGTTTAAACGGTCTCCAGCTTGGCTGTTTTGGTGGATGAGAGAAGATTTTCAGCCTGATACAGATTAAATCAGAACGCAGAAGCGGTCTGATAAAACAGAATTTGCCTGGCGGCAGTAGCGCGGTGGTCCCACCTGACCCCATGCCGAACTCAGAAGTGAAACGCCGTAGCGCCGATGGTAGTGTGGGGTCTCCCCATGCGAGAGTAGGGAACTGCCAGGCATCAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACTCTGAAAGGAGGAACTATATCCGGATCGAGATCC

References

[1] Liu, X., Silverman, A. D., Alam, K. K., Iverson, E., Lucks, J. B., Jewett, M. C., and Raman, S. (2020) Design of a Transcriptional Biosensor for the Portable, On-Demand Detection of Cyanuric Acid. ACS Synth. Biol. 9 (1), 84– 94