BBa_K1362444
BBa_K1362444 Version 1 (Component)
GGG linker with RGK (extein)

RGK
BBa_K1362434 Version 1 (Component)
RGK (extein)

RGD-His-St
BBa_K133036 Version 1 (Component)
RGD-His affinity tag-Stop

R6K
BBa_J61001 Version 1 (Component)
[R6K] Origin of replication

BBa_K1362460
BBa_K1362460 Version 1 (Component)
RGK (extein)

BBa_K1362466
BBa_K1362466 Version 1 (Component)
Intein protease extein

Intein
BBa_K300004 Version 1 (Component)
Engineered pH-inducible intein (codon optimized for E. coli) - internal domain

CWE
BBa_K1362433 Version 1 (Component)
CWE (extein)

Intein
BBa_K1483003 Version 1 (Component)
Ssp GyrB Split Intein

Intein
BBa_K1947002 Version 1 (Component)
Intein

NpuDnaE(N)
BBa_K1362400 Version 1 (Component)
NpuDnaE N-Intein cloning piece

G-His6(N)
BBa_K1362430 Version 1 (Component)
C-terminal His6 (N-Intein)

C2HP/RGE
BBa_K1442107 Version 1 (Component)
C2HP Testing Module (reverse GFP)

NpuDnaE(C)
BBa_K1362401 Version 1 (Component)
NpuDnaE C-Intein cloning piece

BBa_K617003
BBa_K617003 Version 1 (Component)
R6K origin of replication

BBa_K1362251
BBa_K1362251 Version 1 (Component)
Intein protease

Intein C-t
BBa_K2075024 Version 1 (Component)
Intein and Intein Linker (C-term)

Intein N-t
BBa_K2075010 Version 1 (Component)
Intein and Intein linker (N-term)

His6
BBa_K1362431 Version 1 (Component)
N-terminal His6 (C-Intein)

BBa_K1362050
BBa_K1362050 Version 1 (Component)
Intein protease - T7RBS-His-SspDnaBC

Intein
BBa_K2075002 Version 1 (Component)
Intein

BBa_K678004
BBa_K678004 Version 1 (Component)
PGK, mammalian promoter

BBa_K133059
BBa_K133059 Version 1 (Component)
RGD

Intein_assisted_Bisection_Mapping
Intein_assisted_Bisection_Mapping_collection Version 1 (Collection)
Split inteins are powerful tools for seamless ligation of synthetic split proteins. Yet, their use remains limited because the already intricate split site identification problem is often complicated by the requirement of extein junction sequences. To address this, we augmented a mini-Mu transposon-based screening approach and devised the intein-assisted bisection mapping (IBM) method. IBM robustly revealed clusters of split sites on five proteins, converting them into AND or NAND logic gates. We further showed that the use of inteins expands functional sequence space for splitting a protein. We also demonstrated the utility of our approach over rational inference of split sites from secondary structure alignment of homologous proteins. Furthermore, the intein inserted at an identified site could be engineered by the transposon again to become partially chemically inducible, and to some extent enabled post-translational tuning on host protein function. Our work offers a generalizable and systematic route towards creating split protein-intein fusions and conditional inteins for protein activity control.

RGE
BBa_K1442102 Version 1 (Component)
Reverse GFP for E.coli

Intein
BBa_K2075025 Version 1 (Component)
Intein N- and C-Term + Linker

BBa_J61026
BBa_J61026 Version 1 (Component)
[FRT][R6K]

BBa_J61030
BBa_J61030 Version 1 (Component)
[OriTr][R6K]

BBa_K1491024
BBa_K1491024 Version 1 (Component)
ER Intein

R6K ori
BBa_K300998 Version 1 (Component)
[R6K] Origin of replication (rev)

BBa_I50050
BBa_I50050 Version 1 (Component)
R6K replication origin

BBa_K1638034
BBa_K1638034 Version 1 (Component)
Sce VMA Intein

BBa_K404203
BBa_K404203 Version 1 (Component)
ViralBrick-453-RGD

BBa_K404212
BBa_K404212 Version 1 (Component)
ViralBrick-587KO-RGD

BBa_K404207
BBa_K404207 Version 1 (Component)
ViralBrick-587-RGD

BBa_J96014
BBa_J96014 Version 1 (Component)
IgK leader peptide

BBa_J61204
BBa_J61204 Version 1 (Component)
[CmR][TnP][OriTr][R6K]

BBa_K1491026
BBa_K1491026 Version 1 (Component)
Dead ER Intein

BBa_K133013
BBa_K133013 Version 1 (Component)
CF215-RGD-Histop

BBa_K133017
BBa_K133017 Version 1 (Component)
mCherry-RGD-Histop

BBa_K133054
BBa_K133054 Version 1 (Component)
UreB-RGD-Histop

SspDnaB(N)
BBa_K1362408 Version 1 (Component)
SspDnaB N-Intein cloning piece

BBa_K112135
BBa_K112135 Version 1 (Component)
Phagemid-attR1-R6K-attR2

CircNpu
BBa_K1362000 Version 1 (Component)
NpuDnaE intein RFC[105] circularization construct

BBa_K1903019
BBa_K1903019 Version 1 (Component)
Intein N SSp DnaB S1

BBa_J61027
BBa_J61027 Version 1 (Component)
[FRT][R6K][CmR][FRT]