Team:Heidelberg/Parts

From 2010.igem.org

(Difference between revisions)
(composite parts)
(composite parts)
Line 94: Line 94:
|F15||CMV_TetO2||[http://partsregistry.org/Part:BBa_K337027 BBa_K337027]|| CMV promoter under control of Tet Operator for regulation of gene expression
|F15||CMV_TetO2||[http://partsregistry.org/Part:BBa_K337027 BBa_K337027]|| CMV promoter under control of Tet Operator for regulation of gene expression
|-
|-
-
|R1||Sv40(rc)/RSV(fw)||[http://partsregistry.org/Part:BBa_K337045 BBa_K337045]||rowspan="5"| bidirectional hybrid promoter
+
|R1||Sv40(rc)/RSV(fw)||[http://partsregistry.org/Part:BBa_K337045 BBa_K337045]||rowspan="4"| bidirectional hybrid promoter
|-
|-
-
|R2||Sv40(rc)/CMV(fw)||[http://partsregistry.org/Part:BBa_K337047 BBa_K337047]
+
|R2||Sv40(rc)/CMV(fw)||[http://partsregistry.org/Part:BBa_K337047 BBa_K337047]||rowspan="1"| synthetic microRNA template part
|-
|-
|R3||RSV(rc)/CMV(fw)||[http://partsregistry.org/Part:BBa_K337048 BBa_K337048]
|R3||RSV(rc)/CMV(fw)||[http://partsregistry.org/Part:BBa_K337048 BBa_K337048]

Revision as of 02:25, 27 October 2010

Parts

We assembled a lot of 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.

basic parts

table 1: Our miBricks. Listed ID refers to internal nomenclature.
IDContentRegistry linkComment
F1RSV fwBBa_K337012-
F2SV40 rcBBa_K337013-
F3RSV rcBBa_K337014-
F4BGH fwBBa_K337001-
F5BGH rcBBa_K337002 leads to insertion of BamHI site
F7microRNA 10HDBBa_K337016 template for creation of shRNA-like miRNA
F8CMV fwBBa_K337018-
F9FRT siteBBa_K337019-
F10hRlucBBa_K337025-
F16TetRBBa_K337028-
F17Luc2BBa_K337030 compatible for insertion of microRNA binding sites
biCMVbiCMVBBa_K337017-

composite parts

table 2: Our composite parts from subsequent assembling steps. Listed ID refers to internal nomenclature.
IDContentRegistry linkPurpose of the cassette
F6luc2_sv40terBBa_K337015 Luc2 reporter gene with terminator referring to SV40 promoter
F15CMV_TetO2BBa_K337027 CMV promoter under control of Tet Operator for regulation of gene expression
R1Sv40(rc)/RSV(fw)BBa_K337045 bidirectional hybrid promoter
R2Sv40(rc)/CMV(fw)BBa_K337047 synthetic microRNA template part
R3RSV(rc)/CMV(fw)BBa_K337048
R4RSV(rc)/CMV_TetO2(fw)BBa_K337050
R5BGH(rc)/shRNA10(rc) synthetic microRNA template
R13BGH(rc)/shRNA10(rc)/sv40(rc)/RSV(fw)BBa_K337021tuning construct cloning into reporter plasmid backbone containing a binding site against synthetic shRNA-like miRNA
R14BGH(rc)/shRNA10(rc)/sv40(rc)/CMV(fw)BBa_K337022
R15BGH(rc)/shRNA10(rc)/RSV(rc)/CMV(fw)BBa_K337023
R16BGH(rc)/shRNA10(rc)/RSV(rc)/CMV(fw)_TetO2(fw)BBa_K337024
R17BGH(rc)/shRNA6(rc)/sv40(rc)/RSV(fw)BBa_K337026
R18BGH(rc)/shRNA6(rc)/sv40(rc)/CMV(fw)BBa_K337029
R19BGH(rc)/shRNA6(rc)/RSV(rc)/CMV(fw)BBa_K337031
R20BGH(rc)/shRNA6(rc)/RSV(rc)/CMV_TetO2(fw)BBa_K337033tuning construct core containing Tet Operator for On-Targeting together with repressor construct
R21Luc2(rc)_sv40(rc)/CMV(fw)BBa_K337034 bidirectional hybrid promoter with Luc2 reference gene under control of SV40 promoter
R22Luc2(rc)_sv40(rc)/RSV(fw)BBa_K337039
R32Kozag_hRluc/BGH(fw)BBa_K337037reference reporter
R33TetR_mut(XhoI/XbaI)BBa_K337039mutagenized TetR including cutting sites to paste miRNA binding sites
T1BGH(rc)_CMV/TetR/BGH(fw)/BGH(rc)BBa_K337041repressor construct for On-Targeting together with tuning construct
T2BGH(rc)_RSV/TetR/BGH(fw)/BGH(rc)BBa_K337043

final constructs

table 3: Our final constructs. Listed ID refers to internal nomenclature.
IDContentRegistry linkName
K1BGH(rc)/shRNA10(rc)/sv40(rc)/RSV/Luc2_sv40/CMV/Kozag_hRluc_BGHBBa_K337032 pSMB_miTuner Plasmid HD1
K2BGH(rc)/shRNA10(rc)/sv40(rc)/CMV/Luc2_sv40/CMV/Kozag_hRluc_BGHBBa_K337035 pSMB_miTuner Plasmid HD2
K3BGH(rc)/shRNA10(rc)/RSV(rc)/CMV/Luc2_sv40/CMV/Kozag_hRluc_BGHBBa_K337036 pSMB_miTuner Plasmid HD3
K4BGH(rc)/shRNA10(rc)/RSV(rc)/CMV_TetO2/Luc2_sv40/CMV/Kozag_hRluc_BGHBBa_K337038 pSMB_miTuner Plasmid HD4
K5BGH(rc)/shRNA10(rc)/sv40(rc)/RSV/Luc2_sv40/RSV/Kozag_hRluc_BGHBBa_K337040 pSMB_miTuner Plasmid HD5
K6BGH(rc)/shRNA10(rc)/sv40(rc)/CMV/RSV/Luc2_sv40/RSV/Kozag_hRluc_BGHBBa_K337042 pSMB_miTuner Plasmid HD6
K7BGH(rc)/shRNA10(rc)/RSV(rc)/CMV/Luc2_sv40/RSV/Kozag_hRluc_BGHBBa_K337044 pSMB_miTuner Plasmid HD7
K8BGH(rc)/shRNA10(rc)/RSV(rc)/CMV_TetO2/Luc2_sv40/RSV/Kozag_hRluc_BGHBBa_K337046 pSMB_miTuner Plasmid HD8
miMsv40ter(rc)/eBFP(rc)/biCMV/eGFP(fw)/sv40ter(fw)BBa_K337049 pSMB_miMeasure

binding site patterns

table 4: Binding site patterns generated by miraPCR.
IDContentRegistry link
1.3hsa-miR-122 Binding site pattern (3BS)BBa_K337000
1Ahsa-miR-122 Binding site pattern (4BS)BBa_K337003
1-5hsa-miR-122 Binding site pattern (2BS)BBa_K337004
3.7hsa-miR-122 Binding site pattern (2BS - additional 10bp Spacer)BBa_K337005
1.5hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12BBa_K337006
1.8hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12BBa_K337007
3.1hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12BBa_K337008
4.5hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12BBa_K337009
4.6hsa-miR-122 Binding site pattern (2BS) with randomized nt9-12BBa_K337010
mir221-10L2hsa-miR-221 Binding site pattern (2BS)BBa_K337010