Difference between revisions of "Team:BOKU-Vienna/Part Collection"
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<h2>Part Collection overview</h2> | <h2>Part Collection overview</h2> | ||
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| − | <p style="text-align:justify;">For our project the Golden Gate Assembly was used to create the plasmids, which were later transformed into <i>E. coli</i>. The system is based on type IIs restriction enzymes. These cut the DNA outside of their recognition site and create DNA overhangs called fusion sites, which can consist of any nucleotides (shown in figure | + | <p style="text-align:justify;">For our project the Golden Gate Assembly was used to create the plasmids, which were later transformed into <i>E. coli</i>. The system is based on type IIs restriction enzymes. These cut the DNA outside of their recognition site and create DNA overhangs called fusion sites, which can consist of any nucleotides (shown in figure 1). As a result, seamless junctions can be created by designing compatible overhangs. The restriction and ligation event of the desired fragments result in the loss of the original restriction sites. Therefore, it is possible to perform the reaction in a one pot restriction – ligation reaction. </p> |
| − | <p style="text-align:justify;">A hierarchical cloning system was used which contains three levels of complexity, which are shown in figure | + | |
| + | <p style="text-align:justify;">A hierarchical cloning system was used which contains three levels of complexity, which are shown in figure 2. Level 1 called BB1 contains the basic donor plasmids containing genetic building blocks like promotors, genes of interests and terminators. At Level 2 called BB2 a promotor, a gene of interest and a terminator are combined to a transcription unit. The level with the highest complexity is level 3 by combining different transcription units to a multigene construct. At level 3 called BB3 up to 7 BB2 can be combined.</p> | ||
Revision as of 15:05, 31 October 2017
Part Collection
Part Collection overview
For our project the Golden Gate Assembly was used to create the plasmids, which were later transformed into E. coli. The system is based on type IIs restriction enzymes. These cut the DNA outside of their recognition site and create DNA overhangs called fusion sites, which can consist of any nucleotides (shown in figure 1). As a result, seamless junctions can be created by designing compatible overhangs. The restriction and ligation event of the desired fragments result in the loss of the original restriction sites. Therefore, it is possible to perform the reaction in a one pot restriction – ligation reaction.
A hierarchical cloning system was used which contains three levels of complexity, which are shown in figure 2. Level 1 called BB1 contains the basic donor plasmids containing genetic building blocks like promotors, genes of interests and terminators. At Level 2 called BB2 a promotor, a gene of interest and a terminator are combined to a transcription unit. The level with the highest complexity is level 3 by combining different transcription units to a multigene construct. At level 3 called BB3 up to 7 BB2 can be combined.
BB1
• BB1_01Cas9_NLS_FS_23 • BB1_02
cycT_Fs_34 • BB1_03
J23101_FS_12 • BB1_04
pTDH3_FS_34
• BB1_05
B1001_FS_34 • BB1_06
J23105_FS_12 • BB1_07
eGFP_FS_23 • BB1_08
pTEF1_FS_12 • BB1_09
FLPe_FS_23 • BB1_10
repA101ts_FS_23 • BB1_11
MMLV_RT_FS_23 • BB1_12
pBAD_RBS_FS_12 • BB1_13
GFP+PBS1_FS_23 • BB1_14
pGal-syn-v1_FS_12 • BB1_15
J23101_FS_12 • BB1_16
Ura3sgRNA_FS_23 • BB1_17
BB1_17_Leu2sgRNA_FS_23 • BB1_18
AmpR_frameshift_FS_23 • BB1_19
beta_gam_FS_23 • BB1_20
GFP+AlaBS_FS_23 • BB1_21
GFP+ArgBS_FS_23 • BB1_22
GFP+GluBS_FS_23 • BB1_23
GFP+GlyBS_FS_23 • BB1_24
GFP+IleBS_FS_23 • BB1_25
GFP+LeuBS_FS_23 • BB1_26
pGAL_syn-v2_kozak_FS_12 • BB1_27
pBAD+RBS+AraC_FS_12 • BB1_30
pGal_syn_kozak_igem_std_FS_23 • BB1_31
GFP_Improve • BB1_32
GFPuv_FS_23
BB2
• BB2_01J23101_GFP_B1001_AB • BB2_02
pTEF1_FLPe_cycterm_EF • BB2_03
pSNR52_Hefe-Primer1_sup4T_CD • BB2_04
pSNR52_Hefe-Primer2_sup4T_DE • BB2_05
J23101_GFP+PBS1_B1001_AB • BB2_06
J23105_repA101ts_B1001_DE • BB2_07
pBAD_Cas9_B1001_BC • BB2_08
pBAD_RT_FLPe_B1001_BC • BB2_09
Ura3-H1_AB • BB2_10
Ura3-H2_FG • BB2_11
Leu2-H1_AB • BB2_12
Leu2-H2_CD • BB2_13
pGAL-synv1_RT_cycT_BC • BB2_14
J23105_GFP_B1001_AB • BB2_15
J23105_GFP+PBS1_B1001_AB • BB2_16
pGal-syn-v1_GFP_cyc_AB • BB2_17
ori_pSC101(Ts) • BB2_18
pBAD_RT_B1001_BC • BB2_19
J23105_Primer1+HDV_B1001_CD • BB2_20
J23105_AmpR_frameshift_T1001_AB • BB2_21
pBAD_beta_gam_BC • BB2_23
pTEF1_URAsgRNA_cycT • BB2_24
pTEF1_LEUsgRNA_cycT • BB2_25
J23105_GFP+AlaBS_B1001_AB • BB2_26
J23105_GFP+ArgBS_B1001_AB • BB2_27
J23105_GFP+GluBS_B1001_AB • BB2_28
J23105_GFP+GlyBS_B1001_AB • BB2_29
J23105_GFP+IleBS_B1001_AB • BB2_30
J23105_GFP+LeuBS_B1001_AB • BB2_31
PC+AraC_B1001_AB • BB2_32
PC+AraC_B1001_CD • BB2_33
PC+AraC_B1001_DE • BB2_34
J23105_FRT_GFP_B1001_AB • BB2_35
J23105_RNDM_GFP_B1001_AB • BB2_36
pBad_RT+bet+gam_B1001_BC • BB2_37
J23105_GFP+FRT+PBS1_B1001_AB • BB2_38
pGAL-syn-v2_RT_cycT_BC • BB2_39
pGAL-syn-v2_GFP_cycT_AB • BB2_40
pTDH3_GFP_cycT_AB • BB2_41
pBAD+RBS+AraC_Cas9_B1001_BC • BB2_42
J23105_RNDM_GFP_B1001_AB • BB2_43
J23105_ATG_FRT_GFP_B1001_AB • BB2_44
J23105_Primer2+HDV_B1001_CD • BB2_45
J23105_GFP+PBS2_B1001_AB • BB2_46
J23105_GFP+AlaBSv2_B1001_AB • BB2_47
J23105_GFP+GluBSv2_B1001_AB • BB2_48
J23105_GFP+GluBSv2_B1001_AB • BB2_49
J23105_GFP+GlyBSv2_B1001_AB • BB2_50
J23105_GFP+IleBSv2_B1001_AB • BB2_51
AraC+pBAD_GFP+AlaBSv2_B1001_AB • BB2_52
AraC+pBAD_GFP+ArgBSv2_B1001_AB • BB2_53
AraC+pBAD_GFP+GluBSv2_B1001_AB • BB2_54
AraC+pBAD_GFP+GlyBSv2_B1001_AB • BB2_55
AraC+pBAD_GFP+IleBSv2_B1001_AB • BB2_56
J32105_RT_B1001_BC • BB2_57
AraC+pBAD_RT_B1001_BC • BB2_58
AraC+pBAD_GFP+PBS2_B1001_AB • BB2_59
AraC+pBAD+GFP_B1001_AB
BB3
• BB3_01 • BB3_02 • BB3_03 • BB3_04 • BB3_05 • BB3_06 • BB3_07 • BB3_08 • BB3_09 • BB3_10 • BB3_11 • BB3_12 • BB3_13 • BB3_14 • BB3_15 • BB3_16 • BB3_17 • BB3_18 • BB3_19 • BB3_20 • BB3_21 • BB3_22 • BB3_23 • BB3_24 • BB3_25 • BB3_26 • BB3_27 • BB3_28 • BB3_29 • BB3_30 • BB3_31 • BB3_32 • BB3_33 • BB3_34 • BB3_35 • BB3_36 • BB3_37 • BB3_38 • BB3_39 • BB3_40 • BB3_41 • BB3_42 • BB3_43 • BB3_45 • BB3_46 • BB3_47 • BB3_48 • BB3_49 • BB3_55 • BB3_56 • BB3_57 • BB3_58
