pVLT35

Broad-host-range bacterial cloning vector with the pUC18/19 MCS and a spectinomycin/streptomycin resistance gene.
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SalI (9527) XbaI (9521) BamHI (9515) SmaI (9512) KpnI - TspMI - XmaI (9510) Acc65I (9506) SacI (9504) Eco53kI (9502) EcoRI (9494) lac operator MauBI (9219) AclI (9102) MluI (8762) ApaI (8559) PspOMI (8555) HpaI (8260) BsmBI - Esp3I (8147) BspQI - SapI (7954) FspAI (7483) EcoNI (7333) PfoI * (7193) SgrAI (5245) PstI - SbfI (9537) BsaBI * (224) XmnI (338) EagI (377) BsaI (468) PvuI (1231) NgoMIV (1499) NaeI (1501) SfiI (2385) BstZ17I (3760) PasI (4766) pVLT35 9575 bp
SalI  (9527)
1 site
G T C G A C C A G C T G
XbaI  (9521)
1 site
T C T A G A A G A T C T
BamHI  (9515)
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.
SmaI  (9512)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (9510)
1 site
G G T A C C C C A T G G
TspMI  (9510)
1 site
C C C G G G G G G C C C
XmaI  (9510)
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.
Acc65I  (9506)
1 site
G G T A C C C C A T G G
SacI  (9504)
1 site
G A G C T C C T C G A G
Eco53kI  (9502)
1 site
G A G C T C C T C G A G
EcoRI  (9494)
1 site
G A A T T C C T T A A G
MauBI  (9219)
1 site
C G C G C G C G G C G C G C G C
AclI  (9102)
1 site
A A C G T T T T G C A A
MluI  (8762)
1 site
A C G C G T T G C G C A
ApaI  (8559)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (8555)
1 site
G G G C C C C C C G G G
HpaI  (8260)
1 site
G T T A A C C A A T T G
BsmBI  (8147)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (8147)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BspQI  (7954)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (7954)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
FspAI  (7483)
1 site
R T G C G C A Y Y A C G C G T R
EcoNI  (7333)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PfoI  (7193)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SgrAI  (5245)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PstI  (9537)
1 site
C T G C A G G A C G T C
SbfI  (9537)
1 site
C C T G C A G G G G A C G T C C
BsaBI  (224)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
XmnI  (338)
1 site
G A A N N N N T T C C T T N N N N A A G
EagI  (377)
1 site
C G G C C G G C C G G C
BsaI  (468)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
PvuI  (1231)
1 site
C G A T C G G C T A G C
NgoMIV  (1499)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (1501)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
SfiI  (2385)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BstZ17I  (3760)
1 site
G T A T A C C A T A T G
PasI  (4766)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
lacI
8035 .. 9117  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
8035 .. 9117  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
RSF1010 RepB
4599 .. 5570  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepB
4599 .. 5570  =  972 bp
323 amino acids  =  35.9 kDa
Product: replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepC
6907 .. 7758  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepC
6907 .. 7758  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
SmR
852 .. 1643  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
SmR
852 .. 1643  =  792 bp
263 amino acids  =  29.3 kDa
Product: aminoglycoside adenylyltransferase (Murphy, 1985)
confers resistance to spectinomycin and streptomycin
RSF1010 oriV
2537 .. 2931  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
RSF1010 oriV
2537 .. 2931  =  395 bp
replication origin of the broad-host-range plasmid RSF1010; requires the RSF1010 RepA/B/C proteins for replication (Scholz et al., 1989)
T4 terminators
1 .. 116  =  116 bp
T4 transcription termination signals
T4 terminators
1 .. 116  =  116 bp
T4 transcription termination signals
T4 terminators
1903 .. 2018  =  116 bp
T4 transcription termination signals
T4 terminators
1903 .. 2018  =  116 bp
T4 transcription termination signals
RSF1010 oriT
3273 .. 3360  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
3273 .. 3360  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
lacIq promoter
9118 .. 9195  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
9118 .. 9195  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
9494 .. 9550  =  57 bp
pUC18/19 multiple cloning site
MCS
9494 .. 9550  =  57 bp
pUC18/19 multiple cloning site
tac promoter
9425 .. 9453  =  29 bp
3 segments
   Segment 1:  -35  
   9425 .. 9430  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9425 .. 9453  =  29 bp
3 segments
   Segment 2:  
   9431 .. 9446  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9425 .. 9453  =  29 bp
3 segments
   Segment 3:  -10  
   9447 .. 9453  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9425 .. 9453  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
lac operator
9461 .. 9477  =  17 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
9461 .. 9477  =  17 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).
RSF1010 RepA
6081 .. 6920  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 RepA
6081 .. 6920  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  2737 .. 3165  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  3283 .. 3690  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  4189 .. 4602  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  5026 .. 5670  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6808 .. 7758  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  305 .. 613  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  2123 .. 2530  =  408 bp
ORF:  135 amino acids  =  15.0 kDa
ORF:  633 .. 1643  =  1011 bp
ORF:  336 amino acids  =  37.5 kDa
ORF:  3441 .. 5570  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  6081 .. 6920  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  891 .. 1274  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  2958 .. 3242  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  3312 .. 3968  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  4161 .. 4499  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4716 .. 5309  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  6078 .. 6845  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  7248 .. 7589  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
ORF:  122 .. 703  =  582 bp
ORF:  193 amino acids  =  22.1 kDa
ORF:  884 .. 1141  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  1151 .. 1429  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  2159 .. 2404  =  246 bp
ORF:  81 amino acids  =  9.4 kDa
ORF:  3608 .. 3838  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  4505 .. 4858  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  6332 .. 6697  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  8015 .. 8278  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8867 .. 9184  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  352 .. 594  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  2167 .. 2397  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  7141 .. 7479  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  8035 .. 8994  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Download pVLT35.dna file

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