Regulatory

Part:BBa_J23108

Designed by: John Anderson   Group: iGEM2006_Berkeley   (2006-08-17)

constitutive promoter family member For video explanation on this promoter family and its use, see this.

BerkiGEM2006-PromotersEppendorfs.jpg
BerkiGEM2006-Promoters.jpg

 Variant RFP (au)
 J23112           1
 J23103           17
 J23113           21
 J23109           106
 J23117           162
 J23114           256
 J23115           387
 J23116           396
 J23105           623
 J23110           844
 J23107           908
 J23106           1185
 J23108           1303
 J23118           1429
 J23111           1487
 J23101           1791
 J23104           1831
 J23102           2179
 J23100           2547
PBca1020-r0040.jpg

Constitutive promoter family
Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library. J23119 is the "consensus" promoter sequence and the strongest member of the family. All parts except J23119 are present in plasmid J61002. Part J23119 is present in pSB1A2. This places the RFP downstream of the promoter. Reported activities of the promoters are given as the relative fluorescence of these plasmids in strain TG1 grown in LB media to saturation. See part BBa_J61002 for details on their use.

These promoter parts can be used to tune the expression level of constitutively expressed parts. The NheI and AvrII restriction sites present within these promoter parts make them a scaffold for further modification. JCAraw


OUC-China developed part BBa_K2314212 from this part. 5'UTR will be transcribed and can be used as a regulatory element to adjust the translation process. We chose J23108 and added the 5'UTR sequence (UTRrpsT) thereafter. We found that 6h fluorescence intensity increased by more than 1.5- fold. For more information about our new part, please click here:http://parts.igem.org/Part:BBa_K2314212

UTRrpsT enhances protein expression by structure at the 5’UTR.
The enhancement of UTRrpsT may be related to its secondary structure, as shown by the website (http://rna.tbi.univie.ac.at/).
FI/Abs600: J23106 vs J23106+UTR




Manchester 2017 used this part to create part MediumPromoter_PduD(1-20)_mCherry (BBa_K2213007). This promoter was combined with PduD(1-20) to create a tag with medium expression levels. The mCherry tagged PduD(1-20) localisation tag displayed higher fluorescence levels under the medium promoter as compared to under low strength (BBa_K2213006) promoter, and similar fluorescence levels to the high strength (BBa_K2213008) promoter. This result was unexpected and should be taken into consideration when choosing promoters.

More information can be found here: http://parts.igem.org/Part:BBa_K2213007

TagExpression500p.jpg


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/ecoli
//direction/forward
//promoter/anderson
//regulation/constitutive
//rnap/prokaryote/ecoli/sigma70
Parameters
negative_regulators
positive_regulators