Download biGBac enables rapid gene assembly for the expression

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A
B
PmeI
A C
pBIG2ab
loxP
Tn7L
pBIG2ab
(5,668 bp)
GentaR
AmpR
ATAACTTCGT ATAGCATACA TTATACGAAG TTATCTGGCC TAGGTTAATT AAACGCTCTA
PmeI
LoxP
TGGTCTAAAG TTTAAACTGG ATACTATTGC ACGTTTCAAT CAGGAGATCC GAACCAGATA
Tn7R
ori
PacI
Step2 linker A
CamR
Step2 linker C
AGTGAAATCT AGTTCCAAAC TATTTTGTCA TTTTTAATTT TCGTATTAGC TTACGACGCT
PmeI
Tn7L
A D
loxP
Tn7L
pBIG2abc
(5,668 bp)
GentaR
Tn7R
ori
AmpR
CamR
PmeI
A E
loxP
Tn7L
pBIG2abcd
(5,668 bp)
GentaR
Tn7R
ori
AmpR
CamR
PmeI
A F
loxP
Tn7L
pBIG2abcde
(5,668 bp)
GentaR
Tn7R
ori
AmpR
CamR
Fig. S4. pBIG2 vectors. (A) Schematic representation of pBIG2 vectors. pBIG2 vectors are maintained with Chloramphenicol (CamR resistance gene) or Ampicillin (AmpR resistance gene). For the selection in the second assembly step, Chloramphenicol is used. Stocks of linearized pBIG2 cloning vectors are generated
by PmeI digestion. This results in linear vector backbone with linker sequences on both ends. The four pBIG2 vectors contain linker sequence A on one end and
differ only in the linker sequence on the other side (C, D, E, or F as indicated). All biGBac vectors (pLIB, pBIG1, pBIG2) contain Tn7 elements (Tn7L, Tn7R) and a
Gentamicin resistance gene (GentaR) for generation of baculoviruses and a LoxP site for compatibility with Multibac donor plasmids. (B) DNA sequence of
pBIG2ab shown from the LoxP site to Tn7L element. The positions of linker sequences A and C and the restriction sites PmeI and PacI are shown.
Weissmann et al. www.pnas.org/cgi/content/short/1604935113
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