Plasmid backbones/Version 5/Features

1. A complete BioBrick® cloning site for easy cloning and assembly of BioBrick® parts.
2. A ccdB positive selection marker within the BioBrick® cloning site. The ccdB gene ensures that when assembling two BioBrick® parts together, the uncut plasmid vector is not transformed. However, inclusion of the ccdB gene means that these vectors must be propagated in a ccdB tolerant strain, such as E. coli strain DB3.1 (BBa_V1005).
3. A low or medium copy replication origin to reduce consumption of cellular resources during device or system operation.
4. Forward and reverse terminators both upstream and downstream of the BioBrick® cloning site to insulate the vector from read-through transcription originating in the cloned BioBrick® device or system and to insulate the cloned BioBrick® system from the vector.
5. Stop codons in all reading frames to prevent inadvertent translation into or out of the BioBrick cloning site.
6. Primer binding sites for the standard BioBrick® verification primers VF2 (BBa_G00100) and VR (BBa_G00101). These primers are located for convenient sequencing and [http://openwetware.org/wiki/Colony_PCR screening by colony PCR] of cloned BioBrick® devices and systems.