Part:BBa_K1668011

_NOTOC__ mCherry The mCherryis an improved part from BBa_J06702 in Part Registry, composed of RBSBBa_B0034, reporter mCherry BBa_J06702 and double terminator B0010 B0012 .

We found out the sequence of mCherry in 2015 kit plate 3(well 15B) has a promoter with it, therefore doesn’t match the information in Part Registry. Then we sequenced mCherry in 2013, which proved to be correct but in a pSB1A2 backbone, and standardized the correct sequence by assemble the part with standard pSB1C3 backbone. 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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization

BACKGROUND

Figure 1 Iformation of mCherry(BBa_J06702) in Part Registry.

Figure 2 Sequencing results of 2015 mCherry in 2015 kit plate 3, well 15B.

Figure 4 Sequencing results of 2012 mCherry<i> in 2012 kit plate 2, well 8E.

Figure 3 Double enzyme digestion of the 2012<i>mCherry <i>(BBa_J06702) in 2012 kit plate 2, well 8E.

We used <i>mCherry (BBa_J06702) as a reporter, which is one registry star and therefore comparably more reliable. According to the message in the Registry, there is no promoter in the part therefore the reporter should not be expressed(figure 1).

But when we transformed the mCherry gene into E.coli DH5α, the E.coli turned out to become red. With the colorless control, we concluded that the mCherry was expressed and sequnced the parts. According to the results of the sequncing, the part has a promoter with it (figure 2).

Then we got the same part from kit plate in 2013 and 2012. At last, we used mCherry of 2012 in backbone pSB1A2, which is a 2k backbone. But after we have cut the backbone with XbaI and SpeI, we found that the backbone was as big as 4k (figure 3). So we had to sequnce the mCherry of 2012, which turned out to be correct (figure 4). And the backbone had an anti-ampicillin gene with it.

Therefore we concluded that the 2012 mCherry is a correct part with the wrong backbone. Now we had assembled the correct mCherry with pSB1C3 and the mCherry functions well in our other devices (device tcdA1, device plu1537 and device plu0840)