Team:SCUT China/Overview
Parts Overview
We have registered 31 basic parts (BBa_ K3100001-BBa_K310003) and 41 composite parts (BBa_K3100100-BBa_K3100140), all of which is BioBrick RFC10 or Type IIS compatible.
BBa_K3100001 to BBa_K3100010 is different strength T7 promoter which is from the research of Amar Ghodasara et al.[1]. Toehold Switch is prokaryotic riboregulators[2], that activate gene expression in response to cognate RNAs( Trigger RNAs) with arbitrary sequences. We have added Toehold Switch A&C and their Trigger DNAs (Toehold Switch B&D is from previous part number range). BBa_K3100100-BBa_K3100139 are composite parts which are the combination of 4 different Triggers and 10 unique T7 promoters. Through specific design of Fusion Sites, these 40 parts will be randomly combined by Golden Gate assembly. Up to 4 different Triggers driven by 10 unique promoters are expressed from one plasmid and finally a pool called VerProS with a storage capacity of 10000 is obtained. VerProS pool can simultaneously optimize up to four genes in a system. Particularly, this versatile library can be applied to fast optimization in different systems without having to build ad hoc libraries, which can greatly reduce manpower and costs.
We have added 4 acid tolerant factors (BBa_K3100017- BBa_K3100020) and have tested their influence on the acid tolerance of E.coli. What’s more, BBa_K3100140 is a composite part combined of 4 acid tolerant factor and 4 Toehold Switch. In our Project, we use the VerProS pool for fine regulation of these 4 acid tolerant factors.
As T7 is the most well-known inducible promoter with high transcriptional strength [3], we want to construct and improve a T7 promoter library which contains more T7 promoters with different strength. Therefore, the library can help our VerProS system to achieve more fine-grained regulation. Site-saturation mutagenesis was conducted to obtain the T7 mutants. BBa_K3100022-BBa_K3100031 is the different strength T7 promoters which we have constructed and improved.
[1] Ghodasara A, Voigt CA. (2017) Balancing gene expression without library construction via a reusable sRNA pool.
[2] Green AA, Silver PA, Collins JJ, Yin P. (2014) Toehold switches: de-novo-designed regulators of gene expression.
[3] Nie, Z., Luo, H., Li, J. et al. Appl Biochem Biotechnol (2019). https://doi.org/10.1007/s12010-019-03113-y
Parts Table
| BBa_K3100001 | Regulatory | T7 Promoter T7-1 | Zheng Yiyuan | 23 |
| BBa_K3100002 | Regulatory | T7 Promoter T7-2 | Zheng Yiyuan | 23 |
| BBa_K3100003 | Regulatory | T7 Promoter T7-3 | Zheng Yiyuan | 23 |
| BBa_K3100004 | Regulatory | T7 Promoter T7-4 | Zheng Yiyuan | 23 |
| BBa_K3100005 | Regulatory | T7 Promoter T7-6 | Zheng Yiyuan | 22 |
| BBa_K3100006 | Regulatory | T7 Promoter T7-8 | Zheng Yiyuan | 23 |
| BBa_K3100007 | Regulatory | T7 Promoter T7-10 | Zheng Yiyuan | 23 |
| BBa_K3100008 | Regulatory | T7 Promoter T7-15 | Zheng Yiyuan | 23 |
| BBa_K3100009 | Regulatory | T7 Promoter T7-24 | Zheng Yiyuan | 23 |
| BBa_K3100010 | Regulatory | T7 Promoter T7-38 | Zheng Yiyuan | 23 |
| BBa_K3100011 | Regulatory | toehold switch A | Zheng Yiyuan | 90 |
| BBa_K3100012 | Regulatory | toehold switch C | Zheng Yiyuan | 90 |
| BBa_K3100013 | Regulatory | Trigger DNA A | Zheng Yiyuan | 129 |
| BBa_K3100014 | Regulatory | Trigger DNA B | Zheng Yiyuan | 129 |
| BBa_K3100015 | Regulatory | Trigger DNA C | Zheng Yiyuan | 65 |
| BBa_K3100016 | Regulatory | Trigger DNA D | Zheng Yiyuan | 65 |
| BBa_K3100017 | Coding | gadB (antiacid gene) | Zheng Yiyuan | 1401 |
| BBa_K3100018 | Coding | gadC (antiacid gene) | Zheng Yiyuan | 1536 |
| BBa_K3100019 | Coding | katA(antiacid gene) | Zheng Yiyuan | 48 |
| BBa_K3100020 | Coding | ybaS(antiacid gene) | Zheng Yiyuan | 933 |
| BBa_K3100021 | Terminator | rrnBT | Zheng Yiyuan | 247 |
| BBa_K3100022 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | 19 |
| BBa_K3100023 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100024 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100025 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100026 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100027 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100028 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100029 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100030 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100031 | Regulatory | T7 promoter variants family member | Zheng Yiyuan | |
| BBa_K3100100 | Regulatory | T7 Promoter T7-1_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100101 | Regulatory | T7 Promoter T7-1_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100102 | Regulatory | T7 Promoter T7-3_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100103 | Regulatory | T7 Promoter T7-4_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100104 | Regulatory | T7 Promoter T7-6_Trigger DNA A_T7 terminater | Zheng Yiyuan | 215 |
| BBa_K3100105 | Regulatory | T7 Promoter T7-8_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100106 | Regulatory | T7 Promoter T7-10_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100107 | Regulatory | T7 Promoter T7-15_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100108 | Regulatory | T7 Promoter T7-24_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100109 | Regulatory | T7 Promoter T7-38_Trigger DNA A_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100110 | Regulatory | T7 Promoter T7-1_Trigger DNA B_T7 terminater | Zheng Yiyuan | 283 |
| BBa_K3100111 | Regulatory | T7 Promoter T7-2_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100112 | Regulatory | T7 Promoter T7-3_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100113 | Regulatory | T7 Promoter T7-4_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100114 | Regulatory | T7 Promoter T7-6_Trigger DNA B_T7 terminater | Zheng Yiyuan | 215 |
| BBa_K3100115 | Regulatory | T7 Promoter T7-8_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100116 | Regulatory | T7 Promoter T7-10_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100117 | Regulatory | T7 Promoter T7-15_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100118 | Regulatory | T7 Promoter T7-24_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100119 | Regulatory | T7 Promoter T7-38_Trigger DNA B_T7 terminater | Zheng Yiyuan | 216 |
| BBa_K3100120 | Regulatory | T7 Promoter T7-1_Trigger DNA C_T7 terminater | Zheng Yiyuan | 218 |
| BBa_K3100121 | Regulatory | T7 Promoter T7-2_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100122 | Regulatory | T7 Promoter T7-3_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100123 | Regulatory | T7 Promoter T7-4_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100124 | Regulatory | T7 Promoter T7-6_Trigger DNA C_T7 terminater | Zheng Yiyuan | 150 |
| BBa_K3100125 | Regulatory | T7 Promoter T7-8_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100126 | Regulatory | T7 Promoter T7-10_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100127 | Regulatory | T7 Promoter T7-15_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100128 | Regulatory | T7 Promoter T7-24_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100129 | Regulatory | T7 Promoter T7-38_Trigger DNA C_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100130 | Regulatory | T7 Promoter T7-1_Trigger DNA D_T7 terminater | Zheng Yiyuan | 218 |
| BBa_K3100131 | Regulatory | T7 Promoter T7-2_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100132 | Regulatory | T7 Promoter T7-3_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100133 | Regulatory | T7 Promoter T7-4_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100134 | Regulatory | T7 Promoter T7-6_Trigger DNA D_T7 terminater | Zheng Yiyuan | 150 |
| BBa_K3100135 | Regulatory | T7 Promoter T7-8_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100136 | Regulatory | T7 Promoter T7-10_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100137 | Regulatory | T7 Promoter T7-15_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100138 | Regulatory | T7 Promoter T7-24_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100139 | Regulatory | T7 Promoter T7-38_Trigger DNA D_T7 terminater | Zheng Yiyuan | 151 |
| BBa_K3100140 | Coding | antiacid genes combination | Zheng Yiyuan | 4626 |