http://www.osumicrobiology.org/homepages/artsimovitch/sequences/pIAXXX.seq
RNAP
overexpression vectors are highlighted
|
name |
Description |
AbR |
Seq |
|
pIA140, 141 |
PL-nutL-tR’
|
Amp |
|
|
pIA142, 143 |
T7A1-nutR-tR’ |
Amp |
|
|
pIA144, 145 |
PL-nutL-hisT |
Amp |
|
|
pIA146, 147 |
ClaI-BspEI fragment from pRL574 cloned into
pRL777:T7A1, A29, no terminator in sight |
Amp |
|
|
pIA148, 149 |
ClaI-BspEI fragment from pRL574 cloned into
pRM143: lPR, C135, no
terminator in sight |
Amp |
|
|
pIA150 |
Deletion of residues
938-1040 in b - pRL702 derivative |
Amp |
|
|
pIA160 |
XhoI
site in beta at 998 in pRL702 (= His6-b) |
Amp |
|
|
pIA161 |
2H1 fragment (rpoC N-term.) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA162 |
2H2 fragment (rpoC middle) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA163 |
2H3 fragment (rpoC C-term.) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA164 |
2H4 fragment (rpoB N-term.) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA165 |
2H5 fragment (rpoB middle) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA166 |
2H6 fragment (rpoB C-term.) cloned between BamHI and PstI sites of pGAD424 |
Amp |
|
|
pIA167 |
UUUG tetraloop in pCL102b |
Amp |
|
|
pIA168 |
UUUG tetraloop in pCL102b |
Amp |
|
|
pIA169 |
UGA in place of Cys77
codon of lacZ in pRL602 (his promoter) |
Amp |
|
|
pIA170 |
UGA in place of Cys77
codon of lacZ in pIA12 |
Amp |
|
|
pIA171 |
WT downstream sequence
combined with the A29 complex from pCL102b |
Amp |
|
|
pIA172 |
-10, -9, -8 TTT
substitution in pIA171 |
Amp |
|
|
pIA173 |
-10, -9, -8 TTT
substitution combined with GG insertion in the hairpin in pIA171 |
Amp |
|
|
pIA174 |
deletion of b aa 900-909 in pSJ103; contains extra
mutation Ala875Pro- introduces extra BspEI site |
Amp |
|
|
pIA175 |
G to A substitution at the
pause site in pIA171 |
Amp |
|
|
pIA176 |
b F906A in pIA178 |
Amp |
|
|
pIA177 |
b F934A in pIA178 |
Amp |
|
|
pIA178 |
b S531F XhoI
@998 in pRL702 |
Amp |
|
|
pIA179 |
b K909D/S531F in pIA160 |
Amp |
|
|
pIA180 |
b K909D/D516V in pIA160 |
Amp |
|
|
pIA181 |
b D900-909/S531F in pRL702X998 |
Amp |
|
|
pIA182 |
b D900-909 /D516V in
pRL702X998 |
Amp |
|
|
pIA183 |
b D516V in pRL702X998 |
Amp |
|
|
pIA184 |
b’ 762A, 763A, 764A in pRL663 |
Amp |
|
|
pIA185 |
WT put in pIA171 |
Amp |
|
|
pIA186 |
D put in pIA171 |
Amp |
|
|
pIA187 |
b’ 762A, 763A, 764A in pRL662 |
Amp |
|
|
pIA188 |
his71B in pIA171 |
Amp |
|
|
pIA189 |
his71C in pIA171 |
Amp |
|
|
pIA190 |
his71C+mutant hairpin in
pIA171 |
Amp |
|
|
pIA191 |
l PR in pIA170 |
Amp |
|
|
pIA192 |
b K909A in pIA183 |
Amp |
|
|
pIA193 |
b K909S
in pIA183 |
Amp |
|
|
pIA194 |
b K909G
in pIA183 |
Amp |
|
|
pIA195 |
b K909P
in pIA183 |
Amp |
|
|
pIA196 |
b X29Lys6 in pIA183 |
Amp |
|
|
pIA197 |
b F906A
in pIA183 |
Amp |
|
|
pIA198 |
b F934A
in pIA183 |
Amp |
|
|
pIA199 |
B. subtilis P RNA in pCL185 – makes G16 |
Amp |
|
|
pIA200 |
l N cloned between NdeI and XhoI sites of
pET21b - his6 tag at the 3' end. |
Amp |
|
|
pIA201 |
l N cloned between NdeI and XhoI sites of
pET21b |
Amp |
|
|
pIA202 |
-13 through -9 AATTT
substitution in pIA171 |
Amp |
|
|
pIA203 |
mutation in pIA204 - D of C between HphI and SpeI sites |
Amp |
|
|
pIA204 |
his
perfect hairpin in pIA171 |
Amp |
|
|
pIA205 |
b F934A/S531F in pRW408 |
Amp |
|
|
pIA206 |
b K909D/S531F in pRW408 |
Amp |
|
|
pIA207 |
b D900-909/S531F in pRW408 |
Amp |
|
|
pIA208 |
b K909A/S531F in pRW408 |
Amp |
|
|
pIA209 |
b K909S /S531F in pRW408 |
Amp |
|
|
pIA210 |
b F906A/S531F in pRW408 |
Amp |
|
|
pIA211 |
l to cloned between BglII and BamHI sites of pIA214 |
Amp |
|
|
pIA212 |
trpP constitutive promoter cloned between EcoRI and BglII of pIA175 |
Amp |
|
|
pIA213 |
promoter-less CAT gene
cloned into EcoRV site of pET28a |
Amp |
|
|
pIA214 |
promoter-less CAT gene +
US cloned between XbaI and HindIII sites of pIA212 |
Amp |
|
|
pIA215 |
l to cloned between BglII and BamHI sites of pIA214 |
Amp |
|
|
pIA216 |
l tR' cloned between BglII and XbaI sites of pIA214 |
Amp |
|
|
pIA217 |
T7Te cloned between BglII and XbaI sites of pIA214 |
Amp |
|
|
pIA218 |
perfect hairpin that
extends to the -10 position from the 3’ end in pIA171 |
Amp |
|
|
pIA219 |
perfect hairpin that
extends to the -9 position from the 3’ end in pIA171 |
Amp |
|
|
pIA220 |
perfect hairpin that
extends to the -8 position from the 3’ end in pIA171 |
Amp |
|
|
pIA221 |
perfect hairpin that
extends to the -7 position from the 3’ end in pIA171 |
Amp |
|
|
pIA222 |
b D516V with a silent BsgI site - original isolate from Quick Change |
Amp |
|
|
pIA223 |
b D516V with BsgI site - ClaI to BspEI fragment of pIA222 cloned into
pIA160 |
Amp |
|
|
pIA224 |
Mutant 3' proximal region
(UUU->ccc) in T7 terminator in pCM301 |
Amp |
|
|
pIA225 |
Mutant 3' proximal region
(UCU->cuc) in T7 terminator in pCM301 |
Amp |
|
|
pIA226 |
hisP
signal cloned under control of l PR promoter
between BbsI and SphI sites of pCM301 |
Amp |
|
|
pIA227 |
D of b’ in pRM339 |
Amp |
|
|
pIA228 |
his pause signal cloned under
control of of l PR promoter
between BbsI and SphI of pCM301 |
Amp |
|
|
pIA229 |
his perfect hairpin cloned
under control of of l PR promoter
between BbsI and SphI of pCM301 |
Amp |
|
|
pIA230 |
D of NsiI-MfeI fragment (= cI repressor) in
pIA229 |
Amp |
|
|
pIA231 |
NsiI-HindIII fragment from pIA227 into NsiI-HindIII of pBAD33. |
Cm |
|
|
pIA232 |
PCR 3430+3431 from pRL719
cloned between PstI &EcoRI of pBR322 – new StuI site & bla is deleted |
Tet only |
|
|
pIA233 |
killed XhoI site at 10320 in pRL719 |
Amp |
|
|
pIA234 |
SgfI(T4)
fragment of pRL719 cloned between SmaI
and HincII sites of pBSIIKS- :
reverse |
Amp |
|
|
pIA235 |
SgfI(T4)
fragment of pRL719 cloned between SmaI
and HincII sites of pBSIIKS- |
Amp |
|
|
pIA236 |
NsiI
to StuI fragment (9806 bp) of
pRL719 cloned into NsiI & StuI sites of pIA232 |
Tet |
|
|
pIA237 |
ops
site from rfaS cloned between SpeI and BglII sites of pIA171 |
Amp |
|
|
pIA238 |
rfaH
cloned between NdeI and HindIII sites of pET28a |
Kn |
|
|
pIA239 |
S.cerevisiae rpb2 Zn-finger region cloned between Sse87I
and HindIII sites of pIA178 |
Amp |
|
|
pIA240 |
S.cerevisiae rpb2 Zn-finger region cloned between Sse87I
and HindIII sites of pIA233 |
Amp |
|
|
pIA241 |
Derivative of pCM301:
change in the G26 to remove tandem Us + NheI
site |
Amp |
|
|
pIA242 |
b' XbaI
@103 in pIA235 - recloned SseI to MluI fragment from pIA243 into pIA235 |
Amp |
|
|
pIA243 |
b' XbaI
@103 in pIA235 - original isolate from QuikChange mutagenesis |
Amp |
|
|
pIA244 |
WT NusG in pET33 between NdeI and HindIII His6 tag + HMK site at the N-terminus |
Kn |
|
|
pIA245 |
NusG in pET21b between NdeI and HindIII - has a frameshift at the 3’ end + His6 tag |
Amp |
|
|
pIA246 |
WT NusG in pET21b between NdeI and HindIII |
Amp |
|
|
pIA247 |
WT NusG in pET28a between NdeI and HindIII: His6
tag at the N-terminus |
Kn |
|
|
pIA248 |
His6-NusG in
the Ptrc plasmid - NcoI-HindIII fragment of pIA247 cloned into
pRW308 |
Amp |
|
|
pIA249 |
His6-RfaH in
the Ptrc plasmid - NcoI-HindIII fragment of pIA238 cloned into
pRW308 |
Amp |
|
|
pIA250 |
137AT in B. subtilis P RNA – made from pIA199 by
QuikChange mutagenesis |
Amp |
|
|
pIA251 |
ops
site from pheP cloned between SpeI
and BglII sites of pIA171 |
Amp |
|
|
pIA252 |
137AT in B. subtilis P RNA – made from pCP240 by
QuikChange mutagenesis |
Amp |
|
|
pIA253 |
T8 in pCM301 between BbsI
and BamHI |
Amp |
|
|
pIA254 |
T9 in pCM301 between BbsI and BamHI |
Amp |
|
|
pIA255 |
his
terminator in pIA241. PCR 3495-3071 cloned between NheI and BamHI sites of
pIA241 |
Amp |
|
|
pIA256 |
E.coli topA gene fragment under T7A1+A29 - random isolate from cloning PCR
fragment 3483-3484 from genomic DNA into SpeI and BglII sites of pIA171 |
Amp |
|
|
pIA257 |
E.coli fucO gene fragment under T7A1+A29 - random isolate from cloning PCR
fragment 3483-3484 from genomic DNA into SpeI
and BglII sites of pIA171 |
Amp |
|
|
pIA258 |
137AT and 225 TA in B. subtilis P RNA – made from pIA250 by
Quick change mutagenesis |
Amp |
|
|
pIA259 |
137AT and 225TA in B. subtilis P RNA – made from pIA252 by
Quick change mutagenesis |
Amp |
|
|
pIA260 |
ops
from rfaS cloned between SpeI and BglII sites of pIA228 |
Amp |
|
|
pIA261 |
trpPoc
cloned in fron of lacZ: PCR
3932+3933 cloned between HindIII
and EcoRI of pIA12 |
Amp |
|
|
pIA262 |
Substitution of the 9 nt of the ops site in pIA260: GGCGGTAGCGTG to GGCaacgtgcac at 1198-1206 |
Amp |
|
|
pIA263 |
ops
+ T7 terminator from pAR1707 cloned between SpeI and BglII of
pIA228 |
Amp |
|
|
pIA264 |
ops
+ T3 terminator from pCPG T3 cloned between SpeI and BglII of pIA228 |
Amp |
|
|
pIA265 |
ops
+ P14 terminator from pCPG P14 cloned between SpeI and BglII of
pIA228 |
Amp |
|
|
pIA266 |
ops
+ rrnBT1 terminator from pCPG rrnBT1 cloned between SpeI and BglII of pIA228 |
Amp |
|
|
pIA267 |
l tr1 region from l DNA cloned between XmaI
and XbaI of pIA263 |
Amp |
|
|
pIA268 |
ops
site from rfaS cloned between SpeI and BglII sites of pIA171; SpeI
site recreated |
Amp |
|
|
pIA269 |
G to T substitution at nt
2 of the ops site in pIA268 |
Amp |
|
|
pIA270 |
NdeI-HindIII RfaH fragment from pIA249
cloned into pET33-His6-HMK
site at the N-terminus |
Kn |
|
|
pIA271 |
hisP region from pIA171
cloned between SmaI and SphI of pIA263 |
Amp |
|
|
pIA272 |
ops
- terminator fragment cloned between SpeI
and BglII sites of pIA228 (sequence
265-42); should have been P14 but is not |
Amp |
|
|
pIA273 |
GGC to TAA @ 1932-1934 in
pIA260 |
Amp |
|
|
pIA274 |
ops mutant: AACCTTCAC @ 1935-1943 in pIA260 |
Amp |
|
|
pIA275 |
Derivative of pIA263 – SmaI to SphI fragment deleted |
Amp |
|
|
pIA276 |
Derivative of pIA263 – SmaI to SphI fragment deleted, 2 bp deletion relative to 275 |
Amp |
|
|
pIA277 |
Deletion of BamHI fragment in pIA267 - ops site gone |
Amp |
|
|
pIA278 |
RfaH C9S/C28S in pIA238;
SDM |
Kn |
|
|
pIA279 |
RfaH S3C in pIA238; SDM |
Kn |
|
|
pIA280 |
RfaH S173C in pIA238; SDM |
Kn |
|
|
pIA281 |
rpoA
in pGET3A; HindIII site in α is gone - quikchange with oligos
3653+3654; original isolate |
Amp |
|
|
pIA282 |
rpoA
in pET21; HindIII site in α is
gone - quikchange with oligos 3653+3654; original isolate |
Amp |
|
|
pIA283 |
MluI
in pIA171; makes G33; quikchange with oligos 3675 and 3676 |
Amp |
|
|
pIA284 |
MluI
in pIA171; makes G33; quikchange with oligos 3675 and 3676 |
Amp |
|
|
pIA285 |
rfaQ ops in pIA283; just 3 walking steps. Cassette 3677+3678 between MluI and BglII sites of pIA283 |
Amp |
|
|
pIA286 |
XbaI
-HindIII fragment from pIA281
recloned into pGET3A |
Amp |
|
|
pIA287 |
XbaI
-HindIII fragment from pIA281
recloned into pETα |
Amp |
|
|
pIA288 |
Remade pIA174 by quick
change to get rid of Pro875 mutation – original
isolate |
Amp |
|
|
pIA289 |
BspEI-XhoI from pIA288 recloned into pIA178 |
Amp |
|
|
pIA290 |
BamHI-rbs-NcoI linker cloned into BamHI
site of pRL738 |
Amp |
|
|
pIA291 |
BamHI-rbs-NcoI linker cloned into BamHI
site of pRL738 – linker is slightly different from pIA290 |
Amp |
|
|
pIA292 |
NcoI-SseI fragment from pRW408 cloned into
pIA290 |
Amp |
|
|
pIA293 |
NcoI-SseI fragment from pRL702 cloned into
pIA290 |
Amp |
|
|
pIA294 |
b' D943-1130 in pRL663;
original isolate from QuikChange |
Amp |
|
|
pIA295 |
b D226-350 in pIA160;
original isolate from QuikChange with oligos 3593+3694 |
Amp |
|
|
pIA296 |
NcoI-SseI fragment from pIA176 (b S531F/F906A) cloned into pIA291 |
Amp |
|
|
pIA297 |
NcoI-SseI fragment from pIA177 (b S531F/F934A) cloned into pIA291 |
Amp |
|
|
pIA298 |
NcoI-SseI fragment from pIA289 (b S531F/D900-909) cloned into
pIA291 |
Amp |
|
|
pIA299 |
BsmI
to HindIII fragment of pRL736
cloned into pIA292 = T7P-α-β-b’-His6 |
Amp |
|
|
pIA300 |
T7P-α-His6-β[S531F]-β': NcoI-SseI fragment from pIA178 cloned into pIA292 |
Amp |
|
|
pIA301 |
b' D943-1130 in pRL663,
recloned from pIA294; sequenced |
Amp |
|
|
pIA302 |
b D226-350 recloned into
pIA178, sequenced |
Amp |
|
|
pIA303 |
NcoI-SseI fragment from pIA205 cloned into
pIA299 |
Amp |
|
|
pIA304 |
NcoI-SseI fragment from pIA210 cloned into
pIA299 |
Amp |
|
|
pIA305 |
NcoI-SseI fragment from pRW225 cloned into
pIA299 |
Amp |
|
|
pIA306 |
NcoI-SseI fragment from pIA207 cloned into
pIA299 |
Amp |
|
|
pIA307 |
SacI-NcoI deletion in pIA306 |
Amp |
|
|
pIA308 |
b D226-350/D 938-1040/S531F |
Amp |
|
|
pIA309 |
Linker from pIA292 (PCR
product with 3739 and 3740) cloned between BamHI and NcoI sites of
pIA293 |
Amp |
|
|
pIA310 |
Oligo linker (3737+3738)
cloned between BamHI and NcoI sites of pIA293 |
Amp |
|
|
pIA311 |
b 938-1040/S531F |
Amp |
|
|
pIA312 |
T7P-α-β-b’-CBP. Chitin
tag from pTYB3 (3741+3742) digested with SalI
and XhoI & cloned between XhoI sites of pIA299 |
Amp |
|
|
pIA313 |
l PR-B. subtilis P RNA. BamHI
–SphI fragment of pIA199 cloned
into pIA263 |
Amp |
|
|
pIA314 |
T7P-α- His6β[D938-1040]-b’; NcoI-SseI fragment from pIA150 cloned into
pIA309 |
Amp |
|
|
pIA315 |
T7P-α-His6β[S531F/D226-350]-b’; NcoI-SseI fragment from pIA302 cloned into
pIA309 |
Amp |
|
|
pIA316 |
T7P-α-His6β[S531F/D226-350/D938-1040]-b’; NcoI-SseI fragment from pIA308 cloned into pIA309 |
Amp |
|
|
pIA317 |
RfaH C9S in pIA238 |
Kn |
|
|
pIA318 |
RfaH C28S in pIA238 |
Kn |
|
|
pIA319 |
b D226-350 in pIA160 (rifS) |
Amp |
|
|
pIA320 |
b D226-350/D938-1040 in pIA160 (rifS) |
Amp |
|
|
pIA321 |
T7P-α-β-b’[D943-1130]His6; BsmI to HindIII
fragment of pIA301 cloned into pIA299 |
Amp |
|
|
pIA322 |
extension of the 3'
proximal region to 13 nt in pIA171 |
Amp |
|
|
pIA323 |
T7P-α-His6β[F906A]-b’-CBP: NcoI-SdaI fragment from
pIA176 cloned into pIA312 |
Amp |
|
|
pIA324 |
T7P-α-His6β[F934A]-b’-CBP: NcoI-SdaI fragment from
pIA177 cloned into pIA312 |
Amp |
|
|
pIA325 |
T7P-α-His6β[D900-909]-b’-CBP: NcoI-SdaI fragment from
pIA289 cloned into pIA312 |
Amp |
|
|
pIA326 |
extension of the 3'
proximal region to 12 nt in pIA171 - A inserted at -9 |
Amp |
|
|
pIA327 |
extension of the 3'
proximal region to 12 nt in pIA171 - G inserted at -9 |
Amp |
|
|
pIA328 |
extension of the 3'
proximal region to 12 nt in pIA171 - T inserted at -9 |
Amp |
|
|
pIA329 |
T7P-α-His6β[S531F/D226-350]-b’-CBP: NcoI-SdaI fragment from
pIA302 cloned into pIA312 |
Amp |
|
|
pIA330 |
T7P-α-His6β[S531F/D226-350/D983-1040]-b’-CBP: NcoI-SdaI fragment from
pIA308 cloned into pIA312 |
Amp |
|
|
pIA331 |
T7P-α-β-b’[D943-1130]-CBP: BsmI
to XhoI fragment of pIA301 cloned
into pIA312 |
Amp |
|
|
pIA332 |
T7P-α-His6β[S531F]-b’-CBP: NcoI-SdaI fragment from
pIA178 cloned into pIA312 |
Amp |
|
|
pIA333 |
T7P-α-His6β-b’-CBP: NcoI-SdaI fragment from
pIA160 cloned into pIA312 |
Amp |
|
|
pIA334 |
T7P-α-His6β[S531F/D938-1040]-b’-CBP: NcoI-SdaI fragment from pIA311 cloned into
pIA312 |
Amp |
|
|
pIA335 |
RfaH C153S in pIA278 – a single C-terminal Cys |
Kn |
|
|
pIA336 |
T7P-α-His6β[S531F/D226-350]-b’[D943-1130]-CBP: NcoI-SseI fragment from
pIA302 cloned into pIA331 |
Amp |
|
|
pIA337 |
SalI
to KpnI fragment of pIA329 cloned
between XhoI and KpnI sites of Litmus28 |
Amp |
|
|
pIA338 |
SpeI
site in the intergenic region in pIA337 |
Amp |
|
|
pIA339 |
silent NheI site @ b’23/24 + SpeI site in the intergenic region (in pIA337) |
Amp |
|
|
pIA340 |
recloned SseI to EcoRI fragment from pIA339 back into pIA337 |
Amp |
|
|
pIA341 |
SseI
- BsmI from pIA340 recloned into
pIA312; silent silent NheI site @ b’23/24 + SpeI site in intergenic region |
Amp |
|
|
pIA342 |
B. subtilis b fragment from BspEI to F934 (in E. coli) cloned into pIA177 - partial XhoI + BspEI |
Amp |
|
|
pIA343 |
ApaI
to DraIII fragment (large) from
pIA312 cloned into pET28: T7P-α-β-b’-CBP in KnR vector |
Kn |
|
|
pIA344 |
NcoI
to SseI fragment from pIA342 cloned
into pIA312 |
Amp |
|
|
pIA345 |
Long hairpin stem + 12 nt
spacer region |
Amp |
|
|
pIA346 |
ops
site - kps terminator fragment (PCR
3852+3853) cloned between EcoRI and
BamHI sites of pIA12 |
Amp |
|
|
pIA347 |
additional sites BspEI and BglII in pIA285 |
Amp |
|
|
pIA348 |
ops
P – ops P tandem: oligo cassette
3813+3817 cloned between BspEI and XbaI of pIA347 |
Amp |
|
|
pIA349 |
ops
P – his P tandem: PCR 3812-645
cloned between BspEI and XbaI of pIA347 |
Amp |
|
|
pIA350 |
10 nt 3’ proximal region
variant of pIA171 |
Amp |
|
|
pIA351 |
extended hairpin + 11 nt
3'-proximal region: PCR 3818-645 cloned between SpeI and Pst sites of
pIA345 |
Amp |
|
|
pIA352 |
T7P-α-His6β[S531F]-b’[D943-1130]-CBP: NcoI-SseI fragment from
pIA178 cloned into pIA331 |
Amp |
|
|
pIA353 |
ops
+ terminator fragment from kpsF-kpsE
(PCR 3852+3853) cloned between SpeI
and SphI of pIA188 |
Amp |
|
|
pIA354 |
lac
operator in pIA260 between 2000 and 2021 |
Amp |
|
|
pIA355 |
Mutant lacO in pIA260 between 2000
and 2021; substitution A to T at 2009 |
Amp |
|
|
pIA356 |
Mutant lacO operator in pIA260
between 2000 and 2021; 1 bp deletion at 2003 |
Amp |
|
|
pIA357 |
C-less cassette after rfaQ ops site in pIA171 |
Amp |
|
|
pIA358 |
T7P-α[His6]-β-b’-CBP: PCR 3393-3847 from
pIA287 cloned between XbaI and NcoI sites of pIA312 |
Amp |
|
|
pIA359 |
G-less fragment from
pRL540 cloned between SpeI and SphI sites of pIA188 |
Amp |
|
|
pIA360 |
T7P-α[D235-His6]-β- |