Part:BBa_K3365016:Design
pBAD upstream of eGFP with inhibition unit containing lure2
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1053
Illegal XhoI site found at 1062 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 74
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 56
Design Notes
The restriction cutting sites are added to the fragment during PCR for easy ligation to the backbone.
Source
The ligation of BBa_K3365003, BBa_K3365054 and BBa_K3365002 by Overlap PCR.
References
[1] Lu Y, Xue J, Deng T, Zhou X, Yu K, Deng L, Huang M, Yi X, Liang M, Wang Y, Shen H, Tong R, Wang W, Li L, Song J, Li J, Su X, Ding Z, Gong Y, Zhu J, Wang Y, Zou B, Zhang Y, Li Y, Zhou L, Liu Y, Yu M, Wang Y, Zhang X, Yin L, Xia X, Zeng Y, Zhou Q, Ying B, Chen C, Wei Y, Li W, Mok T. Safety and feasibility of CRISPR-edited T cells in patients with refractory non-small-cell lung cancer. Nat Med. 2020 May;26(5):732-740. [2] Guzman LM, Belin D, Carson MJ, Beckwith J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol. 1995 Jul;177(14):4121-30. [3] Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA. Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell. 2013 Feb 28;152(5):1173-83. [4] Vigouroux A, Bikard D. CRISPR Tools To Control Gene Expression in Bacteria. Microbiol Mol Biol Rev. 2020 Apr 1;84(2):e00077-19.