BBa_J119300
1
BBa_J119300
Part for inserting modified D-Dogs for Golden Gate Assembly using BsgI
2013-05-07T11:00:00Z
2015-08-31T04:08:27Z
This is a place holder part only. It has no functionality, thus upon successfully replacing this part with a functional modified C-Dog, expression of mRFP will occur.
This part is a modification of the J100091 part that introduces a modified, non-functional, C-Dog placeholder that when digested with BsgI will allow for insertion of a C-Dog with nested BsgI or BpmI sites. This now allows for the treatment of the C-Dog part as a swappable library tool.
The design of this part is to allow the C-Dog library of parts to become "swappable" in conjunction with optimizing a metabolic network/circuit.
true
false
_613_
0
10385
9
Discontinued
false
The C-Dog parts share a -3 NT overlap with the subsequent coding sequence, making them difficult to swap as part of a library
false
Caleb J. Carr
annotation2217176
1
BsaI cuts left
range2217176
1
28
33
annotation2217175
1
BsaI sticky end left
range2217175
1
23
26
annotation2217183
1
Stop for BD18 leader
range2217183
1
223
225
annotation2217184
1
Start for RFP
range2217184
1
225
227
annotation2217177
1
TT B0014
range2217177
1
34
128
annotation2217187
1
BBa suffix
range2217187
1
906
926
annotation2217181
1
leader RBS
range2217181
1
157
165
annotation2217174
1
BBa Prefix
range2217174
1
1
22
annotation2217188
1
BBa_J100091
range2217188
1
1
926
annotation2217180
1
BD18 bicistron
range2217180
1
140
227
annotation2217178
1
BsaI cuts right
range2217178
1
129
134
annotation2217179
1
BsaI sticky end right
range2217179
1
136
139
annotation2217185
1
RFP E1010
range2217185
1
225
905
annotation2217186
1
Double Stop for E1010
range2217186
1
900
905
annotation2217182
1
RBS for RFP
range2217182
1
210
218
BBa_J100091
1
BBa_J100091
For Testing New Promoters via Golden Gate Assembly
2012-08-19T11:00:00Z
2015-08-31T04:08:22Z
This part was engineered by Todd Eckdahl and Malcolm Campbell. They devised this as part of a national effort to have undergraduates contribute to a registry of functional promoters (RFP; [http://gcat.davidson.edu/RFP/ Registry of Functional Promoters]). There is a very similar part called J119044 [[Part:BBa_J119044]]. The only difference between these two parts is that J00091 uses E1010 [[Part:BBa_E1010]] version of RFP while [[Part:BBa_J119044]] uses an E. coli-optimized set of codons and was produced by GeneArt.
The construct allows for the cloning and testing of new promoter sequences. It is a destination vector for Golden Gate Assembly using BsaI and Ligase, based on part [[Part:BBa_J100028]]. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsaI sites that produce the 4 nt overhangs required for assembly (see [[Part:BBa_J119022]]). Assembly replaces the double terminator in the destination vector with the new promoter. Upon assembly, a functional new promoter will be expected to cause RFP expression. The destination vector also incorporates the BD18 bicistronic translational junction (see [[Part:BBa_J119024]]) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.
false
true
_578_
0
201
61
Not in stock
true
The major question is what plasmid to use. Since both [[Part:BBa_J119044]] and [[Part:BBa_J100091]] both contain a transcriptional terminator (TT), this may lead to recombination if more than two TTs are in the same plasmid. We have some evidence that [[Part:BBa_J119044]] can undergo some recombination when cloned into the plasmid pSB1A8 [[Part:BBa_J119043]]. Therefore, we recommend [[Part:BBa_J119044]] and [[Part:BBa_J100091]] be cloned into a modified version of pSB1A2 that has the Bsa I site removed from the ampicillin gene. We call this modified plasmid pSB1A2 BR for Bsa I Removed.
false
Malcolm Campbell and Todd Eckdahl
annotation2180194
1
leader RBS
range2180194
1
157
165
annotation2180200
1
BBa suffix
range2180200
1
906
926
annotation2180188
1
BsaI sticky end left
range2180188
1
23
26
annotation2180198
1
RFP E1010
range2180198
1
225
905
annotation2180192
1
BsaI sticky end right
range2180192
1
136
139
annotation2330898
1
PCR Rev Primer
range2330898
1
232
251
annotation2180195
1
RBS for RFP
range2180195
1
210
218
annotation2180196
1
Stop for BD18 leader
range2180196
1
223
225
annotation2180190
1
TT B0014
range2180190
1
34
128
annotation2180193
1
BD18 bicistron
range2180193
1
140
227
annotation2180187
1
BBa Prefix
range2180187
1
1
22
annotation2180191
1
BsaI cuts right
range2180191
1
129
134
annotation2180197
1
Start for RFP
range2180197
1
225
227
annotation2180199
1
Double Stop for E1010
range2180199
1
900
905
annotation2180189
1
BsaI cuts left
range2180189
1
28
33
annotation2330897
1
PCR For Primer
range2330897
1
1
20
BBa_J100091_sequence
1
gaattcgcggccgcttctagagcgactgagacctcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattatttggtctcagcgggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcgacggagcgtttctaatggcttcctccgaagatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagattacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataatactagtagcggccgctgcag
BBa_J119300_sequence
1
gaattcgcggccgcttctagagcgactgagacctcacactggctcaccttcgggtgggcctttctgcgtttatatactagagagagaatataaaaagccagattattaatccggcttttttattatttggtctcagcgggggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcgacggagcgtttctaatggcttcctccgaagatgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagatggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagatggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagattacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataatactagtagcggccgctgcag
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z