pQE-TriSystem-7

Vector for expressing untagged proteins in insect and mammalian cells using the QIAgenes system.

Sequence Author: Qiagen

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AanI (3559) AanI (3002) AanI (320) AanI (409) NcoI (2331) EcoRV (2340) TspMI - XmaI (2343) SmaI (2345) BamHI (2350) EcoRI (2356) BglII (2364) AscI (2372) PstI - SbfI (2382) BstBI (2387) HindIII (2390) PaeR7I - XhoI (2399) pQE-TriSystem-7 5250 bp
AanI  (3559)
4 sites
T T A T A A A A T A T T
AanI  (3002)
4 sites
T T A T A A A A T A T T
AanI  (320)
4 sites
T T A T A A A A T A T T
AanI  (409)
4 sites
T T A T A A A A T A T T
NcoI  (2331)
1 site
C C A T G G G G T A C C
EcoRV  (2340)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
TspMI  (2343)
1 site
C C C G G G G G G C C C
XmaI  (2343)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2345)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (2350)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRI  (2356)
1 site
G A A T T C C T T A A G
BglII  (2364)
1 site
A G A T C T T C T A G A
AscI  (2372)
1 site
G G C G C G C C C C G C G C G G
PstI  (2382)
1 site
C T G C A G G A C G T C
SbfI  (2382)
1 site
C C T G C A G G G G A C G T C C
BstBI  (2387)
1 site
T T C G A A A A G C T T
HindIII  (2390)
1 site
A A G C T T T T C G A A
PaeR7I  (2399)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2399)
1 site
C T C G A G G A G C T C
Exon 1
1727 .. 1819  =  93 bp
Exon 1
1727 .. 1819  =  93 bp
Intron
1820 .. 2081  =  262 bp
Intron
1820 .. 2081  =  262 bp
Exon 2
2082 .. 2634  =  553 bp
Exon 2
2082 .. 2634  =  553 bp
AmpR
4328 .. 5188  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4328 .. 5119  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4328 .. 5188  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5120 .. 5188  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4328 .. 5188  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (ORF1629)
2813 .. 3518  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
2813 .. 3518  =  706 bp
contains part of ORF1629
ori
3728 .. 4315  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3728 .. 4315  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
1138 .. 1443  =  306 bp
human cytomegalovirus (CMV) immediate early enhancer
CMV enhancer
1138 .. 1443  =  306 bp
human cytomegalovirus (CMV) immediate early enhancer
chicken β-actin promoter
1449 .. 1726  =  278 bp
chicken β-actin promoter
1449 .. 1726  =  278 bp
T7 terminator
2753 .. 2800  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2753 .. 2800  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF1629
2930 .. 3518  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
2930 .. 3518  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
rabbit globin terminator
2632 .. 2743  =  112 bp
rabbit globin terminator
2632 .. 2743  =  112 bp
p10 promoter
2209 .. 2318  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
2209 .. 2318  =  110 bp
baculovirus promoter for expression in insect cells
MCS
2331 .. 2404  =  74 bp
multiple cloning site
MCS
2331 .. 2404  =  74 bp
multiple cloning site
lac operator
2169 .. 2193  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
2169 .. 2193  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
8xHis
2432 .. 2455  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2432 .. 2455  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
siRNA target site
2407 .. 2425  =  19 bp
siRNA target site
2407 .. 2425  =  19 bp
stop codons
2456 .. 2466  =  11 bp
stop codons in all three reading frames
stop codons
2456 .. 2466  =  11 bp
stop codons in all three reading frames
RBS
2321 .. 2326  =  6 bp
ribosome binding site
RBS
2321 .. 2326  =  6 bp
ribosome binding site
poly(A) signal
2610 .. 2615  =  6 bp
poly(A) signal
2610 .. 2615  =  6 bp
ATG
2333 .. 2335  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2333 .. 2335  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  1315 .. 1704  =  390 bp
ORF:  129 amino acids  =  13.9 kDa
ORF:  1547 .. 1798  =  252 bp
ORF:  83 amino acids  =  8.7 kDa
ORF:  4458 .. 4724  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5238 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  387 .. 617  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  1671 .. 2060  =  390 bp
ORF:  129 amino acids  =  13.3 kDa
ORF:  2300 .. 2596  =  297 bp
ORF:  98 amino acids  =  11.2 kDa
ORF:  2930 .. 3547  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  4328 .. 5188  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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