Difference between revisions of "Part:BBa K1614002"
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===Usage and Biology===
 
===Usage and Biology===
 
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The BioBrick Standard as described by BBF RFC 10 enables the assembly of multiple DNA fragments or modules called parts. Its key advantage is that the assembly of several parts can be performed by the repetitive use of a simple protocol using only a small set of relatively cheap methods and enzymes. Also the limitation to only few restriction enzymes lowered the likelihood for the presence of cut sites in the genes of interest. Yet the use of traditional restriction cloning presents significant disadvantages when it comes to DNA elements where the cloning product has to be seamless, like for protein fusions, and even has to be sequence specific base-by-base. This is the case if the RNA transcript of the sequence is responsible for the function of the part and not a translated protein. As every base may determine the tertiary structure and thereby behavior of the functional RNA, there are no “wobble bases” that allow the easy substitution of a base.
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The BioBrick Standard as described by BBF RFC 10 enables the assembly of multiple DNA fragments or modules called parts. Its key advantage is that the assembly of several parts can be performed by the repetitive use of a simple protocol using only a small set of relatively cheap methods and enzymes. Also the limitation to only few restriction enzymes lowered the likelihood for the presence of cut sites in the genes of interest. Yet the use of traditional restriction cloning presents significant disadvantages when it comes to DNA elements where the cloning product has to be seamless, like for protein fusions, and even has to be sequence specific base-by-base. This is the case if the RNA transcript of the sequence is responsible for the function of the part and not a translated protein. As every base may determine the tertiary structure and thereby behavior of the functional RNA, there are no “wobble bases” that allow the easy substitution of a base (Figure 1).
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[[File:HeidelbergPWiGEM2015_RFC.png|thumb|'''Fig. 1.'''General mechanism of the RFC which is based on BioBrick BBa_K1614002]]
 
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Revision as of 15:23, 20 September 2015

transcription cassette of BBF RFC 110

Unit for RFC 110 assembly, clonable in RFC 10 vectors like pSB1C3.

Usage and Biology

The BioBrick Standard as described by BBF RFC 10 enables the assembly of multiple DNA fragments or modules called parts. Its key advantage is that the assembly of several parts can be performed by the repetitive use of a simple protocol using only a small set of relatively cheap methods and enzymes. Also the limitation to only few restriction enzymes lowered the likelihood for the presence of cut sites in the genes of interest. Yet the use of traditional restriction cloning presents significant disadvantages when it comes to DNA elements where the cloning product has to be seamless, like for protein fusions, and even has to be sequence specific base-by-base. This is the case if the RNA transcript of the sequence is responsible for the function of the part and not a translated protein. As every base may determine the tertiary structure and thereby behavior of the functional RNA, there are no “wobble bases” that allow the easy substitution of a base (Figure 1).

Fig. 1.General mechanism of the RFC which is based on BioBrick BBa_K1614002

This part is the core of the RFC we propose for the generation of functional RNA. It comprises a cassette for generating well-defined end of the RNA-sequence, thereby guaranteeing predictable and reproducible functionality.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 32
    Illegal NgoMIV site found at 61
  • 1000
    COMPATIBLE WITH RFC[1000]