Team:Freiburg/Notebook/10 June

From 2011.igem.org

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Enzymes used for digestion:
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|RBS (S4-S6)
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|Vector (S7)
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Revision as of 12:45, 8 July 2011

Contents

??.Labday

Miniprep

Investigators: Jakob, Julia


Comments: Trafo was performed on 8th of june and plates were incubated over night at 37°C. Clones were picked to do a miniprep.


Miniprep of parts from 8th of june and DNA concentration was measured with NanoDrop 2000c.

3A-assembly

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

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

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