Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Raik Gruenberg
Date created: 2008-08-10 11:00:00
Date modified: 2015-08-31 04:08:35
pSB1AK3F construction plasmid for protein fusion BioBricks
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_J18902_sequence (Version 1) |
Description
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 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
Notes
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.
Source
constructed from pSB1AK3| Sequence Annotation | Location | Component / Role(s) |
| extended BioBrick suffix extended BioBrick prefix KanR AmpR BBF2 BBVR Replication Origin (MB1) | 1,30 3177,3205 1106,1921 2076,2936 3058,3078 166,185 270,883 | feature/misc sequence_feature feature/misc sequence_feature CDS feature/cds feature/cds CDS primer_binding_site feature/primer_binding primer_binding_site feature/primer_binding sequence_feature feature/misc |