Difference between revisions of "Part:BBa J63010:Design"
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===Design Notes=== | ===Design Notes=== | ||
This plasmid was designed to permit in-frame assembly of protein fusions. Although it contains the standard biobrick ends (EcoRI, XbaI, SpeI, PstI), a part immediately follows the XbaI restriction site and immediately precedes a SpeI site. These changes form a [http://hdl.handle.net/1721.1/32535 revised biobrick assembly strategy]. When two parts in this plasmid are fused in the standard assembly method, six nucleotides (ACTAGA) separate the two parts, thus keeping the coding sequences in-frame. | This plasmid was designed to permit in-frame assembly of protein fusions. Although it contains the standard biobrick ends (EcoRI, XbaI, SpeI, PstI), a part immediately follows the XbaI restriction site and immediately precedes a SpeI site. These changes form a [http://hdl.handle.net/1721.1/32535 revised biobrick assembly strategy]. When two parts in this plasmid are fused in the standard assembly method, six nucleotides (ACTAGA) separate the two parts, thus keeping the coding sequences in-frame. | ||
| − | Like the biobrick standard, parts cannot contain EcoRI, XbaI, SpeI or PstI restriction sites. An additional design criteria is that a part should not begin with the nucleotides TC. If a part did, the XbaI site would be methylated by DamI and thus digestion at this site would be blocked. | + | *Like the biobrick standard, parts cannot contain EcoRI, XbaI, SpeI or PstI restriction sites. |
| + | *An additional design criteria is that a part should not begin with the nucleotides TC. If a part did, the XbaI site would be methylated by DamI and thus digestion at this site would be blocked. | ||
===Source=== | ===Source=== | ||
Revision as of 14:43, 29 October 2006
Protein fusion vector (Silver lab standard)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3246
Illegal XbaI site found at 3261
Illegal SpeI site found at 1
Illegal PstI site found at 15 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3246
Illegal SpeI site found at 1
Illegal PstI site found at 15
Illegal NotI site found at 8
Illegal NotI site found at 3252 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3246 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Illegal EcoRI site found at 3246
Illegal XbaI site found at 3261 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3246
Illegal XbaI site found at 3261
Illegal SpeI site found at 1
Illegal PstI site found at 15
Illegal NgoMIV site found at 2825 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 1393
Illegal SapI site found at 310
Design Notes
This plasmid was designed to permit in-frame assembly of protein fusions. Although it contains the standard biobrick ends (EcoRI, XbaI, SpeI, PstI), a part immediately follows the XbaI restriction site and immediately precedes a SpeI site. These changes form a [http://hdl.handle.net/1721.1/32535 revised biobrick assembly strategy]. When two parts in this plasmid are fused in the standard assembly method, six nucleotides (ACTAGA) separate the two parts, thus keeping the coding sequences in-frame.
- Like the biobrick standard, parts cannot contain EcoRI, XbaI, SpeI or PstI restriction sites.
- An additional design criteria is that a part should not begin with the nucleotides TC. If a part did, the XbaI site would be methylated by DamI and thus digestion at this site would be blocked.
Source
synthetized DNA
References
[http://hdl.handle.net/1721.1/32535 1. Ira Phillips and Pam Silver. "A New Biobrick Assembly Strategy Designed for Facile Protein Engineering." DSpace at MIT].