Parts
We assembled many miBricks (i. e. basic parts) to composite parts that were then used to built up our final constructs. The submitted parts are listed below. We did not categorize the parts according to different subprojects since they are universally applicable or clearly denoted through their names. Some binding site patterns, which were created via [... miraPCR], are listed separately.
Main Measurement Constructs - Engineered
ID | Content | Registry link | Name
|
K3 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV/Luc2_sv40/CMV/Kozag_hRluc_BGH | BBa_K337036 | pSMB_miTuner Plasmid HD3
|
K4 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV_TetO2/Luc2_sv40/CMV/Kozag_hRluc_BGH | BBa_K337038 | pSMB_miTuner Plasmid HD4
|
miM | sv40ter(rc)/eBFP(rc)/biCMV/eGFP(fw)/sv40ter(fw) | BBa_K337049 | pSMB_miMeasure
|
Synthetic Single Binding Sites
(Physical DNA not submitted)
Design | Content | Registry link | Name
|
KD:97% | perfect binding site | BBa_K337052 | shRNA miRhaat
|
KD:69% | imperfect binding site: point mut 11 | BBa_K337053 | shRNA miRhaat
|
KD:28% | imperfect binding site: bulge 16-18 | BBa_K337054 | shRNA miRhaat
|
KD:96% | perfect binding site | BBa_K337055 | miR122
|
KD:64% | imperfect binding site: | BBa_K337056 | miR122
|
KD:24% | imperfect binding site: | BBa_K337057 | miR122
|
Synthetic MicroRNA Binding Site Patterns
ID | Content | Registry link
|
1.3 | hsa-miR-122 Binding site pattern (3BS) | BBa_K337000
|
1A | hsa-miR-122 Binding site pattern (4BS) | BBa_K337003
|
1-5 | hsa-miR-122 Binding site pattern (2BS) | BBa_K337004
|
3.7 | hsa-miR-122 Binding site pattern (2BS - additional 10bp Spacer) | BBa_K337005
|
1.5 | hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12 | BBa_K337006
|
1.8 | hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12 | BBa_K337007
|
3.1 | hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12 | BBa_K337008
|
4.5 | hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12 | BBa_K337009
|
4.6 | hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12 | BBa_K337010
|
mir221-10L2 | hsa-miR-221 Binding site pattern (2BS) | BBa_K337011
|
miTunig Kit - Basic Parts
miTuning Kit - Intermediate Parts 1
ID | Content | Registry link | Purpose of the cassette
|
F6 | luc2_sv40ter | BBa_K337015 | Luc2 reporter gene with terminator referring to SV40 promoter
|
F15 | CMV_TetO2 | BBa_K337027 | CMV promoter under control of Tet Operator for regulation of gene expression
|
R1 | Sv40(rc)/RSV(fw) | BBa_K337045 | bidirectional hybrid promoter
|
R2 | Sv40(rc)/CMV(fw) | BBa_K337047
|
R3 | RSV(rc)/CMV(fw) | BBa_K337048
|
R4 | RSV(rc)/CMV_TetO2(fw) | BBa_K337050
|
R5 | BGH(rc)/shRNA10(rc) | BBa_K337051 | synthetic microRNA template part
|
R13 | BGH(rc)/shRNA10(rc)/sv40(rc)/RSV(fw) | BBa_K337020 | tuning construct cloning into reporter plasmid backbone containing a binding site against synthetic shRNA-like miRNA
|
R14 | BGH(rc)/shRNA10(rc)/sv40(rc)/CMV(fw) | BBa_K337021
|
R15 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV(fw) | BBa_K337022
|
R16 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV(fw)_TetO2(fw) | BBa_K337023
|
R17 | BGH(rc)/shRNA6(rc)/sv40(rc)/RSV(fw) | BBa_K337024
|
R18 | BGH(rc)/shRNA6(rc)/sv40(rc)/CMV(fw) | BBa_K337026
|
R19 | BGH(rc)/shRNA6(rc)/RSV(rc)/CMV(fw) | BBa_K337029
|
R20 | BGH(rc)/shRNA6(rc)/RSV(rc)/CMV_TetO2(fw) | BBa_K337031 | tuning construct core containing Tet Operator for On-Targeting together with repressor construct
|
R21 | Luc2(rc)_sv40(rc)/CMV(fw) | BBa_K337033 | bidirectional hybrid promoter with Luc2 reference gene under control of SV40 promoter
|
R22 | Luc2(rc)_sv40(rc)/RSV(fw) | BBa_K337034
|
R32 | Kozag_hRluc/BGH(fw) | BBa_K337037 | reference reporter
|
R33 | TetR_mut(XhoI/XbaI) | BBa_K337039 | mutagenized TetR including cutting sites to paste miRNA binding sites
|
T1 | BGH(rc)_CMV/TetR/BGH(fw)/BGH(rc) | BBa_K337041 | repressor construct for On-Targeting together with tuning construct
|
T2 | BGH(rc)_RSV/TetR/BGH(fw)/BGH(rc) | BBa_K337043
|
Tuning Kit - Intermediate Parts 2
ID | Content | Registry link | Name
|
K1 | BGH(rc)/shRNA10(rc)/sv40(rc)/RSV/Luc2_sv40/CMV/Kozag_hRluc_BGH | BBa_K337032 | pSMB_miTuner Plasmid HD1
|
K2 | BGH(rc)/shRNA10(rc)/sv40(rc)/CMV/Luc2_sv40/CMV/Kozag_hRluc_BGH | BBa_K337035 | pSMB_miTuner Plasmid HD2
|
K5 | BGH(rc)/shRNA10(rc)/sv40(rc)/RSV/Luc2_sv40/RSV/Kozag_hRluc_BGH | BBa_K337040 | pSMB_miTuner Plasmid HD5
|
K6 | BGH(rc)/shRNA10(rc)/sv40(rc)/CMV/RSV/Luc2_sv40/RSV/Kozag_hRluc_BGH | BBa_K337042 | pSMB_miTuner Plasmid HD6
|
K7 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV/Luc2_sv40/RSV/Kozag_hRluc_BGH | BBa_K337044 | pSMB_miTuner Plasmid HD7
|
K8 | BGH(rc)/shRNA10(rc)/RSV(rc)/CMV_TetO2/Luc2_sv40/RSV/Kozag_hRluc_BGH | BBa_K337046 | pSMB_miTuner Plasmid HD8
|