Part:BBa_K316011

Reverse strand coding XylE

This part contains XylE coding on the 3' strand. In many biological settings there may be read-through caused by upstream elements, however this can often be unidirectional. In settings where readthrough from 5' direction is expected to be much stronger than 3' direction, it may is advantageous to use 3' coding sequence instead of traditional 5'.

For the purposes of characterisation, the part was made on the minus strand to reduce background transcription. Read-through from 5' direction may provide an additional benefit. Low quantities of anti-sence RNA may be produced as the result of 5' readthrough, which may in turn reduce basal expression sometimes seen in LacI systems.

For more information about XylE, it's substrate and spectrophotometric assays, please see BBa_K316003 or our [http://2010.igem.org/Team:Imperial_College_London/Results wiki results section]

Safety

The substrate XylE works on is a chemical called catechol. It is classed as irritant in the EU but as toxic in the USA, as well as being a possible carcinogen. It should therefore be handled with care and proper safety equipment. More information is available on the Material Safety Data Sheet[http://www.sciencelab.com/msds.php?msdsId=9927131 Material Safety Data Sheet].

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Characterisation data was obtained for XylE BBa_K316003. In addition constructs under two different promoters: J23101-XylE BBa_K316004 from E. coli was used to categorise B. subtilis derived Pveg-XylE BBa_K316005. Also GFP-XylE constructs BBa_K316007 were tested to determine the effectiveness of inhibition of XylE activity by attachment of GFP. These are described on our wiki[http://2010.igem.org/Team:Imperial_College_London/Results] and the aforementioned parts pages.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 437
  Illegal NgoMIV site found at 609
  Illegal AgeI site found at 86
  
• 1000
  COMPATIBLE WITH RFC[1000]