pSB1AT3F construction plasmid for protein fusion BioBricks
pSB1AT3F is almost identical to pSB1AT3 but uses the "Freiburg" extension of the standard BioBrick prefix and suffix and features silent mutations in the tetracycline resistance gene. The Freiburg extension adds a new pair of inner restriction enzymes -- NgoMIV (prefix) and AgeI (suffix) -- which allows the in-frame fusion of protein parts. See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats BB formats]. pSB1AT3F is a high copy number plasmid carrying ampicillin and tetracyclin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010. The given sequence starts with the Freiburg BB suffix and ends with the Freiburg BB prefix. The complete vector sequence is therefore obtained from: part + vector
This plasmid was constructed by PCR and InFusion recombination. 1) The pSB1AK3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites 2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites 3) The two overlapping PCR products were recombined using the Clonetech InFusion kit. 4) Two NgoMIV restriction sites in the original pSB1AT3 (within the TcR) were deleted following the QuickChange protocol (from Qiagen). PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AT3. Only insert and flanks have been verified by sequencing. The deletion of NgoMIV sites has been verified by restriction digests.
pSB1A3 with an C-terminal Arginine tag. This particular plasmid utilizes the BioFusion (aka Silver Lab) Standard for BioBrick assembly. Adds 10 Arginine residues to the C-terminus of a Protein (Includes Stop Codon)
pSB1A3 with an N-terminal Arginine tag. This particular plasmid utilizes the BioFusion (aka Silver Lab) Standard for BioBrick assembly. Adds 10 Arginine residues to the N-terminus of a Protein.
pSB1AC3F construction plasmid for protein fusion BioBricks
pSB1AC3F is identical to pSB1AC3 but uses the "Freiburg" extension of the standard BioBrick prefix and suffix. This Freiburg extension adds a new pair of inner restriction enzymes -- NgoMIV (prefix) and AgeI (suffix) -- which allows the in-frame fusion of protein parts. See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats BB formats]. pSB1AC3F is a high copy number plasmid carrying ampicillin and chloramphenicol resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010. The given sequence starts with the Freiburg BB suffix and ends with the Freiburg BB prefix. The complete vector sequence is therefore obtained from: part + vector
This plasmid was constructed by PCR and InFusion recombination. 1) The pSB1AC3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites 2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites 3) The two overlapping PCR products were recombined using the clonetech InFusion kit. All PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AC3. Only insert and flanks have been verified by sequencing.
pSB1AK3F construction plasmid for protein fusion BioBricks
pSB1AK3F is identical to pSB1AK3 but uses the "Freiburg" extension of the standard BioBrick prefix and suffix. This Freiburg extension adds a new pair of inner restriction enzymes -- NgoMIV (prefix) and AgeI (suffix) -- which allows the in-frame fusion of protein parts. See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats BB formats]. pSB1AK3F is a high copy number plasmid carrying ampicillin and kanamycin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010. The given sequence starts with the Freiburg BB suffix and ends with the Freiburg BB prefix. The complete vector sequence is therefore obtained from: part + vector
This plasmid was constructed by PCR and InFusion recombination. 1) The pSB1AK3 vector backbone was linearized by PCR, introducing the NgoMIV / AgeI restriction sites 2) The P1010 insert was amplified by PCR, introducing the NgoMIV / AgeI restriction sites 3) The two overlapping PCR products were recombined using the clonetech InFusion kit. All PCR reactions were performed with a proof-reading polymerase featuring a very low mutation rate. Nevertheless, the vector backbone may contain mutations with respect to pSB1AC3. Only insert and flanks have been verified by sequencing.
Low copy protein fusion vector (Silver lab standard)
Silver Lab alias V0002. This is a modified biobrick vector (fusion brick vector) that contains the specified EcoRI, SpeI, XbaI, and PstI sites. The only difference is that the part sequence immediately follows the XbaI site and immediately precedes the SpeI site. This change allows in-frame fusions of protein coding regions to be made using standard biobricks assembly methods.
If the part starts with the nucleotides tc, then that part-containing plasmid must be grow in a DamI- strain to avoid methylation (and inhibition of digestion) at the XbaI site.
Plasmid with BioBrick 3.0 prefix and suffix
Protein fusion vector (Silver lab standard)
standard biofusion plasmid containing ccdB death gene and chlorenphenicol resistance