Difference between revisions of "Part:BBa K1152013"
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a) Definition of different domain border combinations for T-domain exchanges
 
a) Definition of different domain border combinations for T-domain exchanges

Revision as of 03:00, 5 October 2013

IndC Indigoidine Synthetase device


This part encodes the non-ribosomal peptide synthetase that synthesizes the blue pigment indigoidine. The IndC gene was amplified from P. luminescens laumondii TT01 (DSM15139) and cloned into pSB1C3.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4087
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1467
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2730

Restriction digest of BBa_K1152013 with EcoRI, XbaI, SpeI and PstI


<a href="Heidelberg_IndPD_Fig6.png"><IMG HEIGHT=50 WIDTH=50 SRC="Heidelberg_IndPD_Fig6.png">

a) Definition of different domain border combinations for T-domain exchanges The figure shows a sequence alignment of the indC and bpsA amino acid sequences. The alignment was created using clustalO (http://www.ebi.ac.uk/Tools/msa/clustalo/) with standard parameters. The lines marked A, B and C reflect the borders we used between the A- and the T-domain, whereas those marked, 1, 2, 3 and 4 reflect the borders between the T- and the TE-domain. In total we tried all twelve combinations of a domain border {A, B, C} and a domain border {1, 2, 3, 4}, replacing the sequence inbetween with the respective part of bpsA.

b) E. coli TOP10 co-transformed with modified versions of indC and the PPTase sfp The co-tranformation of the modified indC-(bpsA-T) plasmids described above with a second plasmid coding for the PPTase Sfp shows that only three domain border combinations can be used for exchanging the indC T-domain with the T-domain of bpsA. These are the combinations A1, A2 and C1. We applied combination A2 for further T-domain exchanges.