Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-12 12:03:11
Lac Monomer A w/ 16 bp spacer for ICA
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_K2066011_sequence (Version 1) |
Description
The part is flanked by UNS 2 and UNS 3 as per the WM UNS gibson cloning standard. This part is the LacO, 16BP spacer 'A' monomer for use in ICA to create repeated TetO Arrays of variable length. The monomer can be PCR amplified using UNS 4 and UNS 5 PCR landing pads. Next, the part should cut with BsmBI restriction enzyme to expose sticky ends and used to extend monomer in ICA. Monomer A (this part) can bind to the initiator sequence (available on our wiki) or Monomer C. This part contains the following: UNS 2 (WM standard gibson/amplification primer site) ??? UNS 4 (used to provide orthogonal amplification of monomers alone) ??? BsmBI site ??? sticky end 1 - Lac Repeat ??? 16 bp spacer (taken as first 16 bp from UNS X) ??? BsmBI site ??? Sticky end 2 - UNS 5 (orthogonal amplification in conjunction with UNS 4) ??? UNS 3 (WM standard gibson/amplification primer site).ICA method and sequence design based on Briggs et al., 2012 and its supplement.
Briggs, A. W., Rios, X., Chari, R., Yang, L., Zhang, F., Mali, P., & Church, G. M. (2012). Iterative capped assembly: rapid and scalable synthesis of repeat-module DNA such as TAL effectors from individual monomers. Nucleic acids research, gks624.
The UNS2, UNS3, UNS 4, and UNS 5 sequences are taken from Torella et al. 2013 and are ideal for cloning long sequences of monomers.
Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860.
Notes
Design of monomer part was based off of TALEN monomer design in Briggs et al., 2012.Source
ICA method and sequence design based on Briggs et al., 2012 and its supplement. Briggs, A. W., Rios, X., Chari, R., Yang, L., Zhang, F., Mali, P., & Church, G. M. (2012). Iterative capped assembly: rapid and scalable synthesis of repeat-module DNA such as TAL effectors from individual monomers. Nucleic acids research, gks624. The UNS2, UNS3, UNS 4, and UNS 5 sequences are taken from Torella et al. 2013 and are ideal for cloning long sequences of monomers. Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860.| Access | Instance | Definition |
| public public public public public public public public | BBa_K2066018 BBa_K2066020 BBa_K2066501 BBa_K2066525 BBa_K2066514 BBa_K2066504 BBa_K2066021 BBa_K2066019 | BBa_K2066018 BBa_K2066020 BBa_K2066501 BBa_K2066525 BBa_K2066514 BBa_K2066504 BBa_K2066021 BBa_K2066019 |