Richard Lab:Bio-Brick Formats

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(Biobrick Standard)
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===Biobrick Standard===
===Biobrick Standard===
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This standard uses the typical biobrick assembly enzymes [EcoRI (GAATTC), XbaI (TCTAGA), SpeI (ACTAGT), and PstI (CTGCAG)] as well as NotI sites (GCGGCCGC) flanking the part. There is a slightly different prefix for protein coding parts to put the start codon in a better spot in relation to the RBS.
+
This standard uses the typical biobrick assembly enzymes [EcoRI (GAATTC), XbaI (TCTAGA), SpeI (ACTAGT), and PstI (CTGCAG)] as well as NotI sites (GCGGCCGC) flanking the part. There is a slightly different prefix for protein coding parts to put the start codon in a better spot in relation to the RBS.  During assembly the XbaI site of the "rear part" and the SpeI site of the "front" part eliminate eachother and form a "scar" which forever binds the two parts together.
*Prefix
*Prefix
::*GAATTC'''gcggccgc'''tTCTAGAg
::*GAATTC'''gcggccgc'''tTCTAGAg

Revision as of 16:29, 24 January 2011

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Contents

Introduction

In general the Richard Lab uses the standard Bio-Brick format. However occasionally this format is incompatible with our goals (e.g. making protein fusions) or is unnecessarilly cumbersome (e.g. PCR overlap extension). To have a review on why (and how) we use these sites see the our New Standard Assembly page.

Formats

Biobrick Standard

This standard uses the typical biobrick assembly enzymes [EcoRI (GAATTC), XbaI (TCTAGA), SpeI (ACTAGT), and PstI (CTGCAG)] as well as NotI sites (GCGGCCGC) flanking the part. There is a slightly different prefix for protein coding parts to put the start codon in a better spot in relation to the RBS. During assembly the XbaI site of the "rear part" and the SpeI site of the "front" part eliminate eachother and form a "scar" which forever binds the two parts together.

  • Prefix
  • GAATTCgcggccgctTCTAGAg
  • The EcoRI and XbaI sites are in CAPS while the NotI site is in bold.
  • Protein Coding Prefix
  • GAATTCgcggccgctTCTAG
  • using this prefix the first base of the start codon (i.e. the "A" in "ATG") will complete the XbaI site.
  • Suffix
  • tACTAGTagcggccgCTGCAG

Short Biobrick

Protein Fusions

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