BBa_K886003
1
BBa_K886003
Recombination Device composed by Cre-lox71
2012-09-23T11:00:00Z
2015-05-08T01:13:40Z
We synthesized the lox71 site, then we assembled it downstream the Cre recombinase gene(BBa_J61047).
This device was created to express the Cre recombinase enzyme and a target gene under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR, using primers that will insert a loxP and a lox66 sites flanking the sequence, in order to integrate it in the lox71 recombination site. For this part the lox71 is located downstream the Cre recombinase gene. An alternative construction is submitted in the part BBa_K886001.
Once this device is located under the control of the desired promoter, it will express the Cre recombinase that recognizes the lox sites and inserts the target gene under the control of the same promoter.
The Cre expression levels are critical for the recombination, we created two different devices: BBa_K886002 (Cre-Lox71) and BBa_K886001 (Lox71-Cre) for testing if inserting the target gene either upstream or downstream the Cre gene will affect the expression efficiency of the target gene
false
false
_940_
0
11298
9
It's complicated
false
We used the lox71 submitted by our group BBa_K886000, which corrected the parts BBa_I718017 and BBa_K537020. Those parts are wrong when compared with the published sequence at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC137435/
false
edgar andres ochoa cruz
component2196284
1
BBa_J61047
component2196285
1
BBa_K886000
annotation2196285
1
BBa_K886000
range2196285
1
1046
1079
annotation2196284
1
BBa_J61047
range2196284
1
1
1037
BBa_K886000
1
BBa_K886000
Fixed lox71
2012-09-23T11:00:00Z
2015-05-08T01:13:40Z
primers
Released HQ 2013
The particularity of lox71 is that it has an altered sequence at the end of it's right arm compared to loxP. This sequence variation reduces affinity of the Cre recombinase for the arm.
false
false
_1149_
0
11298
9
In stock
false
Based on the report made by the igem2010 UT-Tokyo team and some papers, the lox71 (BBa_I718017) from the registry was wrong, it had a cg instead a gc in its center sequence that is between the arms.
Apparently, this part was corrected by the iGEM11_WITS_CSIR_SA team (BBa_K537020), but the sequence from this group had the same error. The corrected part from iGEM11_Tokyo_Tech (BBa_K649205) was right but no DNA was available in the registry. So we decided to synthesized it and test it, using the proper sequence described by http://www.ncbi.nlm.nih.gov/pmc/articles/PMC137435/, and we submitted the correct DNA to the Registry.
false
edgar andres ochoa cruz
BBa_J61047
1
Cre
Cre DNA recombinase
2007-02-20T12:00:00Z
2015-08-31T02:03:00Z
bob
bob
false
false
_95_
0
483
95
In stock
false
bob
true
John Anderson
BBa_K886000_sequence
1
taccgttcgtatagcatacattatacgaagttat
BBa_J61047_sequence
1
atgtccaatttactgaccgtacaccaaaatttgcctgcattaccggtcgatgcaacgagtgatgaggttcgcaagaacctgatggacatgttcagggatcgccaggcgttttctgagcatacctggaaaatgcttctgtccgtttgccggtcgtgggcggcatggtgcaagttgaataaccggaaatggtttcccgcagaacctgaagatgttcgcgattatcttctatatcttcaggcgcgcggtctggcagtaaaaactatccagcaacatttgggccagctaaacatgcttcatcgtcggtccgggctgccacgaccaagtgacagcaatgctgtttcactggttatgcggcggatccgaaaagaaaacgttgatgccggtgaacgtgcaaaacaggctctagcgttcgaacgcactgatttcgaccaggttcgttcactcatggaaaatagcgatcgctgccaggatatacgtaatctggcatttctggggattgcttataacaccctgttacgtatagccgaaattgccaggatcagggttaaagatatctcacgtactgacggtgggagaatgttaatccatattggcagaacgaaaacgctggttagcaccgcaggtgtagagaaggcacttagcctgggggtaactaaactggtcgagcgatggatttccgtctctggtgtagctgatgatccgaataactacctgttttgccgggtcagaaaaaatggtgttgccgcgccatctgccaccagccagctatcaactcgcgccctggaagggatttttgaagcaactcatcgattgatttacggcgctaaggatgactctggtcagagatacctggcctggtctggacacagtgcccgtgtcggagccgcgcgagatatggcccgcgctggagtttcaataccggagatcatgcaagctggtggctggaccaatgtaaatattgtcatgaactatatccgtaacctggatagtgaaacaggggcaatggtgcgcctgctggaagatggcgattaagaatt
BBa_K886003_sequence
1
atgtccaatttactgaccgtacaccaaaatttgcctgcattaccggtcgatgcaacgagtgatgaggttcgcaagaacctgatggacatgttcagggatcgccaggcgttttctgagcatacctggaaaatgcttctgtccgtttgccggtcgtgggcggcatggtgcaagttgaataaccggaaatggtttcccgcagaacctgaagatgttcgcgattatcttctatatcttcaggcgcgcggtctggcagtaaaaactatccagcaacatttgggccagctaaacatgcttcatcgtcggtccgggctgccacgaccaagtgacagcaatgctgtttcactggttatgcggcggatccgaaaagaaaacgttgatgccggtgaacgtgcaaaacaggctctagcgttcgaacgcactgatttcgaccaggttcgttcactcatggaaaatagcgatcgctgccaggatatacgtaatctggcatttctggggattgcttataacaccctgttacgtatagccgaaattgccaggatcagggttaaagatatctcacgtactgacggtgggagaatgttaatccatattggcagaacgaaaacgctggttagcaccgcaggtgtagagaaggcacttagcctgggggtaactaaactggtcgagcgatggatttccgtctctggtgtagctgatgatccgaataactacctgttttgccgggtcagaaaaaatggtgttgccgcgccatctgccaccagccagctatcaactcgcgccctggaagggatttttgaagcaactcatcgattgatttacggcgctaaggatgactctggtcagagatacctggcctggtctggacacagtgcccgtgtcggagccgcgcgagatatggcccgcgctggagtttcaataccggagatcatgcaagctggtggctggaccaatgtaaatattgtcatgaactatatccgtaacctggatagtgaaacaggggcaatggtgcgcctgctggaagatggcgattaagaatttactagagtaccgttcgtatagcatacattatacgaagttat
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