Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Tom Knight
Date created: 2006-03-16 12:00:00
Date modified: 2015-05-08 01:13:51
Right mosaic end for Tn5 transpososome
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_M1001_sequence (Version 1) |
Description
Right end of a linear transpososome sequence containing the 19 bp mosaic end in the reverse orientation. The sequence also contains a PvuII cut site which releases the transpososome when cut with PvuII. Constructed compound parts intended for transposition must not contain other PvuII end, or be PCR amplified with blunt end producing enzymes (Pfu, Phusion) at the PvuII cut site between CAG and CTG.This part, and the companion part M1000 provide Biobrick tools to construct Tn5 transpososomes: linear DNA sequences which can be cut in vitro from a plasmid backbone, or created by PCR, combined in vitro with purified Tn5 trasposase protein, electroportated into cells, and finally randomly inserting into the bacterial chromosome.
The parts assembled between the two mosaic ends can be delivered, including antibiotic resistance markers or other functional chromosomal insertions.
Notes
;biobrick left end, TA cloning overhangGTT TCT TCG AAT TCG CGG CCG CTT CTA GAG
;biobrick right end, TA cloning overhang
GTT TCT TCC TGC AGC GGC CGC TAC TAG TA
;mosaic end with cag upstream to insert PvuII cut site
cagCTGTCTCTTATACACATCT
;mosaic end reverse complement with ctg upstream to insert PvuII cut site
AGATGTGTATAAGAGACAGctg
;mosaic left end in biobrick form construction primer F
GTT TCT TCG AAT TCG CGG CCG CTT CTA GAG cagCTGTCTCTTATACACATCT, MEL-F
;mosaic left end in biobrick form construction primer R
GTT TCT TCC TGC AGC GGC CGC TAC TAG TA AGATGTGTATAAGAGACAGctg, MEL-R
;mosaic right end in biobrick form construction primer F
GTT TCT TCG AAT TCG CGG CCG CTT CTA GAG AGATGTGTATAAGAGACAGctg, MER-F
;mosaic right end in biobrick form construction primer R
GTT TCT TCC TGC AGC GGC CGC TAC TAG TA cagCTGTCTCTTATACACATCT, MER-R
Source
Synthesis from overlapped oligos with PCR.| Sequence Annotation | Location | Component / Role(s) |
| PvuII cut site Mosaic end (inverted) | 17,22 1,19 | sequence_feature feature/misc sequence_feature feature/misc |