Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Julie Norville
Date created: 2009-03-25 12:00:00
Date modified: 2015-05-08 01:08:20
A BioScaffold Part (Uses PpiI), similar to J70010 see Part Design Page
| Types | DnaRegion |
| Roles | DNA sequence_feature |
| Sequences | BBa_J70210_sequence (Version 1) |
Description
When placed in a BioBrick assembly and cut with the restriction enzyme PpiI, this part will direct the restriction enzyme to cut outside the BioBrick scarstill in testing, uses PpiI (2) PacI (2) MabI (2), currently in pSB1AK3 vector
See part design page or BBFRFC15, for details about use
This part can be made with the following oligos, which can be annealed and placed into an EcoRI/SpeI cut vector:
TM=54.3 TM=53.8 TM=58.6
EcoRINotI XbaI SpeI
AATTCGCGGCCGCTTCTAGAG| gagagctggttcaccaggtt aattaattaattaaacctggtgaacgtggtctc ta (ctag)
G | at gatc
gaattcgcggccgcttctagag ta ctagtagcggccgctgcag
BioBrick Prefix gagagctggttcaccaggtt aattaattaattaaacctggtgaacgtggtctc BioBrick Suffix
J70010 sequence
Oligos for J70010.2 (3/17/09)
Forward 1.2 (TM=54.3): 5'-AATTCGCGGCCGCTTCTAGAG-3' (this oligo corrects an error from version 1.1)
Forward 2.2 (TM1=53.8, TM2=58.6): 5'-gagagctggttcaccaggttaattaattaattaaacctggtgaacgtggtctcta-3'
Reverse 2.2 (TM1=53.8,54.3): 5'-aacctggtgaaccagctctcCTCTAGAAGCGGCCGCG-3' (This oligo corrects a version 2.1 error)
Reverse 1.2: 5'-ctagtagagaccacgttcaccaggtttaattaattaatt-3'
Obsolete Versions:
Forward 1.1: AATTCGCGGCCGCTACTAGAG (bug)
Forward 2.1: gagagctggttcaccaggttaattaattaattaaacctggtgaacgtggtctcta
Reverse 1.1: CTAGTAGAgaccacgttcaccaggtttaattaattaatt
Reverse 2.1: aacctggtgaaccagctctcCTCTAGTAGCGGCCGCG (bug)
Notes
Same as J70010 except uses AscI instead of PacICan be used to cut at 0 offset from the BioBrick scar on either side, see BBF RFC15
TACTAGAG and TACAGAT represent sequences for BioBrick scars that are created when
two BioBrick parts are fused together.
^s represent restriction enzyme cut sites.
General Function
XXXXX XXXXX^TACTAGAG BioScaffold Part BBa_J70010 TACTAGAT XXXXX^XXXXX
XXXXX^XXXXX ATGATCAC ATGATCTA^XXXXX XXXXX
Function when the right part is a protein part.
XXXXX XXXXX^TACTAGAG BioScaffold Part BBa_J70010 TACTAGAT GXXXX^XXXXX
XXXXX^XXXXX ATGATCAC ATGATCTA^CXXXX XXXXX
What happens after cutting with MabI or PacI and purifying with either gel purification
or a Qiagen PCR cleanup spin column
General Case
XXXXX XXXXX^ ^XXXXX
XXXXX^ ^XXXXX XXXXX
Case when right part is a protein part
XXXXX XXXXX^ ^XXXXX
XXXXX^ ^CXXXX XXXXX
Source
designed| Sequence Annotation | Location | Component / Role(s) |
| PpiI PpiI PpiI PpiI MabI dna AscI AscI | 1,3 9,12 43,46 52,54 4,9 45,51 20,27 28,35 | feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/misc feature/misc sequence_feature sequence_feature feature/dna feature/dna sequence_feature non_covalent_binding_site feature/binding non_covalent_binding_site feature/binding |