Team:Freiburg/Notebook/10 June
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+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/9_June">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 10 June </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/11_June">Next entry</a> | ||
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==Miniprep== | ==Miniprep== | ||
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Miniprep of parts from 8th of june and DNA concentration was measured with NanoDrop 2000c. | Miniprep of parts from 8th of june and DNA concentration was measured with NanoDrop 2000c. | ||
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- | ==3A-assembly== | + | ==3A-assembly: Digestion== |
'''Investigators: Sophie, Theo, Sandra''' | '''Investigators: Sophie, Theo, Sandra''' | ||
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|PstI | |PstI | ||
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- | ==Protein modelling== | + | ==<span style="color:grey;">Protein modelling for the precipitator</span>== |
'''Investigators: Rüdiger''' | '''Investigators: Rüdiger''' |
Latest revision as of 01:18, 22 September 2011
3A-assembly: Digestion
Investigators: Sophie, Theo, Sandra
Digestion of RBS and promotor to create a promotor-RBS library.
sample | name | |
Promotor | BBa_J23104 | S1 |
Promotor | BBa_J23110 | S2 |
Promotor | BBa_J23116 | S3 |
RBS | BBa_B0034 | S4 |
RBS | BBa_B0032 | S5 |
RBS | BBa_B0031 | S6 |
Vector | pSB1C3 | S7 |
Enzymes used for digestion:
enzyme 1 | enzyme 2 | |
promotor (S1-S3) | EcoRI | SpeI |
RBS (S4-S6) | Xbal | PstI |
Vector (S7) | EcoRI | PstI |
Protein modelling for the precipitator
Investigators: Rüdiger
3cvr bacterial ligase
FSELQLNRLNLSSLPDNLPPQ
ITVLEITQNALISLPELPAS
LEYLDACDNRLSTLPELPAS
LKHLDVDNNQLTXLPELPAL
LEYINADNNQLTXLPELPTS
LEVLSVRNNQLTFLPELPES
EIFFRCRENRITHIPENILSLDP
Extracted ideal consensus, by consensus sequence and 3d rational analysis:
LEVLDVSNNQLTSLPDNLPAS
N-Terminal Hagfish 2z66
CPSRCSCSGTEIRCNSKGLTSVPTGIPSSATRLELESNKLQSLPHGVFDK
LTQLTK
Consensus: xLxxLxxLxL
C-Terminal Hagfish 2z62
LKELALDTNQLKSVPDGIFDR
LTSLQKIWLHTNPWDCSCPRIDY
LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
LxxLxLxxNxLx Consensus
Sequences 1-6 sent to I-Tasser for 3D structure prediction: Rational1: LxALHCSxNxLxSLPxxLPxx LxHLACSxNxLxSLPxxLPxx
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS ATRLELESNKLQSLPHGVFDK LTALHKSNNQLTSLPDNLPAS LEHLAVSNNQLTSLPDNLPAS LEALHVSNNQLTSLPDNLPAS LEHLAVSNNQLTSLPDNLPAS LEALHVSNNQLTSLPDNLPAS LEHLAVSNNQLTSLPDNLPAS LKELALDTNQLKSVPDGIFDR LTSLQKIWLHTNPWDCSCPRIDY LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
Rational2: LxxLxCSxNHLHSLPxxLPxx
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS
ATRLELESNKLQSLPHGVFDK
LTQLTKSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LKELALDTNQLKSVPDGIFDR
LTSLQKIWLHTNPWDCSCPRIDY
LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
Rational3:
LxxLxCSxNxLxSLPHHLPxx
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS ATRLELESNKLQSLPHGVFDK LTQLTKSNNQLTSLPHHLPAS LEVLDVSNNQLTSLPHHLPAS LEVLDVSNNQLTSLPHHLPAS LEVLDVSNNQLTSLPHHLPAS LEVLDVSNNQLTSLPHHLPAS LEVLDVSNNQLTSLPHHLPAS LKELALDTNQLKSVPDGIFDR LTSLQKIWLHTNPWDCSCPRIDY LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
Rational4: LxxLxCSxNxLxSLPxHLPHx
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS ATRLELESNKLQSLPHGVFDK LTQLTKSNNQLTSLPDHLPHS LEVLDVSNNQLTSLPDHLPHS LEVLDVSNNQLTSLPDHLPHS LEVLDVSNNQLTSLPDHLPHS LEVLDVSNNQLTSLPDHLPHS LEVLDVSNNQLTSLPDHLPHS LKELALDTNQLKSVPDGIFDR LTSLQKIWLHTNPWDCSCPRIDY LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
Rational5:
LHxLxCSxNxLxSLPxxLPxH
LHVLDVSNNQLTSLPDNLPAH
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS
ATRLELESNKLQSLPHGVFDK
LHQLTKSNNQLTSLPDNLPAH
LHVLDVSNNQLTSLPDNLPAH
LHVLDVSNNQLTSLPDNLPAH
LHVLDVSNNQLTSLPDNLPAH
LHVLDVSNNQLTSLPDNLPAH
LHVLDVSNNQLTSLPDNLPAH
LKELALDTNQLKSVPDGIFDR
LTSLQKIWLHTNPWDCSCPRIDY
LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
Rational6: LHHLxCSxNxLxSLPxxLPxx LHHLDVSNNQLTSLPDNLPAS
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS
ATRLELESNKLQSLPHGVFDK
LHHLTKSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LHHLDVSNNQLTSLPDNLPAS
LKELALDTNQLKSVPDGIFDR
LTSLQKIWLHTNPWDCSCPRIDY
LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP