Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Reshma Shetty
Date created: 2008-03-22 12:00:00
Date modified: 2015-08-31 04:07:42
BioBrick base vector, as synthesized by Codon Devices
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_I51019_sequence (Version 1) |
Description
[[Part:BBa_I51019|BBa_I51019]] is the sequence of the BioBrick base vector as synthesized by Codon Devices, Inc. in Cambridge, MA. The original design of the BioBrick base vector is available as [[Part:BBa_I51000|BBa_I51000]]. [[Part:BBa_I51001|BBa_I51001]] is the sequence of the BioBrick base vector as reported by Codon Devices, Inc. although the true sequence is something like to [[Part:BBa_I51019|BBa_I51019]]. The exact sequence of the provided DNA is difficult to determine. The corrected BioBrick base vector that was successfully constructed from [[Part:BBa_I51019|BBa_I51019]] is [[Part:BBa_I51020|BBa_I51020]].Notes
Synthesis of the BioBrick base vector design ([[Part:BBa_I51000|BBa_I51000]]) was problematic. We encountered two issues while trying to construct the base vector. First, troubleshooting efforts during synthesis compromised the design of our parts: failed attempts to clone the base vector into an E. coli strain intolerant of expression of the toxic protein CcdB led to an unnecessary redesign of the ccdB positive selection marker in the BioBrick base vector. As a result, a revised design for the BioBrick base vector ([[Part:BBa_I51001|BBa_I51001]]) was specified instead. Second, faulty part design adversely impacted the synthesis process: our pUC19-based replication origin design ([[Part:BBa_I50020|BBa_I50020]]) was similarly nonfunctional, so the revised base vector ([[Part:BBa_I51001|BBa_I51001]]) could not be propagated as specified. We eventually determined that the provided DNA for the BioBrick base vector was actually a fusion of two slightly different copies of the base vector: one with a nonfunctional version of the pUC19 origin ([[Part:BBa_I50020|BBa_I50020]]) and one with a functional version of the pUC19 origin ([[Part:BBa_I50022|BBa_I50022]]). We corrected the synthesized BioBrick base vector ([[Part:BBa_I51019|BBa_I51019]]) with molecular biology techniques to make the functional BioBrick base vector [[Part:BBa_I51020|BBa_I51020]] (see Methods). Commercial DNA synthesis processes currently rely on cloning, assembly and propagation of synthesized DNA in E. coli. Difficulties during synthesis stemmed from the inclusion of both a ccdB positive selection marker that is toxic to most E. coli strains and a synthetic origin incapable of supporting plasmid replication of the BioBrick base vector. In general, for parts whose function are incompatible with growth and replication of E. coli, the processes of DNA design and DNA synthesis cannot be easily decoupled.Source
[[Part:BBa_I51001|BBa_I51001]]| Access | Instance | Definition |
| public public public public public public public public public public public public public public public public public public public public public public public public public public public public public public public public public public | BBa_G00001 BBa_B0044 BBa_B0042 BBa_B0054 BBa_G00102 BBa_B0062 BBa_B0045 BBa_P1006 BBa_B0045 BBa_B0053 BBa_G00100 BBa_B0055 BBa_B0042 BBa_B0043 BBa_G00000 BBa_P1016 BBa_I50020 BBa_G00001 BBa_B0044 BBa_B0042 BBa_B0054 BBa_G00102 BBa_B0062 BBa_B0045 BBa_P1006 BBa_B0045 BBa_B0053 BBa_G00100 BBa_B0055 BBa_B0042 BBa_B0043 BBa_G00000 BBa_P1016 BBa_I50022 | Suffix TOPO TSS BBa_B0054 VR BBa_B0062 NheI ampR NheI BBa_B0053 VF2 BBa_B0055 TSS TOPO Prefix ccdB pUC19 ori Suffix TOPO TSS BBa_B0054 VR BBa_B0062 NheI ampR NheI BBa_B0053 VF2 BBa_B0055 TSS TOPO Prefix ccdB pUC19 ori |