Difference between revisions of "Part:BBa J63010:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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transform into TOP10 survival
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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 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.
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+
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.
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===Source===
 
===Source===

Revision as of 14:38, 29 October 2006


Protein fusion vector (Silver lab standard)


Assembly Compatibility:
  • 10
    INCOMPATIBLE 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
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3246
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Illegal EcoRI site found at 3246
    Illegal XbaI site found at 3261
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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 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