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Plasmid Files

pVLT31

Broad-host-range bacterial cloning vector with the pUC18/19 MCS and a tetracycline resistance gene.

 
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pVLT31.dna
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XbaI (9615) SmaI (9606) KpnI - TspMI - XmaI (9604) Acc65I (9600) SacI (9598) Eco53kI (9596) EcoRI (9588) MauBI (9313) MluI (8856) ApaI (8653) PspOMI (8649) HpaI (8354) BsmBI (8241) BspQI - SapI (8048) PfoI * (7287) AcuI (5729) PasI (4860) PstI - SbfI (9631) DraIII (414) NheI (644) BmtI (648) PshAI (1131) EagI (1354) NruI (1389) BsaBI * (1888) SfiI (2479) BstZ17I (3854) pVLT31 9669 bp
XbaI  (9615)
1 site
T C T A G A A G A T C T
SmaI  (9606)
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  (9604)
1 site
G G T A C C C C A T G G
TspMI  (9604)
1 site
C C C G G G G G G C C C
XmaI  (9604)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
Acc65I  (9600)
1 site
G G T A C C C C A T G G
SacI  (9598)
1 site
G A G C T C C T C G A G
Eco53kI  (9596)
1 site
G A G C T C C T C G A G
EcoRI  (9588)
1 site
G A A T T C C T T A A G
MauBI  (9313)
1 site
C G C G C G C G G C G C G C G C
MluI  (8856)
1 site
A C G C G T T G C G C A
ApaI  (8653)
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  (8649)
1 site
G G G C C C C C C G G G
HpaI  (8354)
1 site
G T T A A C C A A T T G
BsmBI  (8241)
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.
BspQI  (8048)
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  (8048)
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.
PfoI  (7287)
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.
AcuI  (5729)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PasI  (4860)
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.
PstI  (9631)
1 site
C T G C A G G A C G T C
SbfI  (9631)
1 site
C C T G C A G G G G A C G T C C
DraIII  (414)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
NheI  (644)
1 site
G C T A G C C G A T C G
BmtI  (648)
1 site
G C T A G C C G A T C G
PshAI  (1131)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
EagI  (1354)
1 site
C G G C C G G C C G G C
NruI  (1389)
1 site
T C G C G A A G C G C T
BsaBI  (1888)
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.
SfiI  (2479)
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  (3854)
1 site
G T A T A C C A T A T G
TcR
501 .. 1691  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
501 .. 1691  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
lacI
8129 .. 9211  =  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
8129 .. 9211  =  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
4693 .. 5664  =  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
4693 .. 5664  =  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
7001 .. 7852  =  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
7001 .. 7852  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriV
2631 .. 3025  =  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
2631 .. 3025  =  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
1997 .. 2112  =  116 bp
T4 transcription termination signals
T4 terminators
1997 .. 2112  =  116 bp
T4 transcription termination signals
RSF1010 oriT
3367 .. 3454  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
3367 .. 3454  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
lacIq promoter
9212 .. 9289  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
9212 .. 9289  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
9588 .. 9644  =  57 bp
pUC18/19 multiple cloning site
MCS
9588 .. 9644  =  57 bp
pUC18/19 multiple cloning site
tac promoter
9519 .. 9547  =  29 bp
   Segment 1:  -35  
   9519 .. 9524  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9519 .. 9547  =  29 bp
   Segment 2:  
   9525 .. 9540  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9519 .. 9547  =  29 bp
   Segment 3:  -10  
   9541 .. 9547  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9519 .. 9547  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
lac operator
9555 .. 9571  =  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
9555 .. 9571  =  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
6175 .. 7014  =  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
6175 .. 7014  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  3535 .. 5664  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  6175 .. 7014  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  674 .. 1057  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  1595 .. 1855  =  261 bp
ORF:  86 amino acids  =  9.7 kDa
ORF:  2831 .. 3259  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  3377 .. 3784  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  4283 .. 4696  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  5120 .. 5764  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6902 .. 7852  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  501 .. 1691  =  1191 bp
ORF:  396 amino acids  =  41.5 kDa
ORF:  2217 .. 2624  =  408 bp
ORF:  135 amino acids  =  15.0 kDa
ORF:  3052 .. 3336  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  3406 .. 4062  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  4255 .. 4593  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4810 .. 5403  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  6172 .. 6939  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  7342 .. 7683  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
ORF:  738 .. 980  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  1038 .. 1496  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  2253 .. 2498  =  246 bp
ORF:  81 amino acids  =  9.4 kDa
ORF:  3702 .. 3932  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  4599 .. 4952  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  6426 .. 6791  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  8109 .. 8372  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  8961 .. 9278  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  854 .. 1204  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  2261 .. 2491  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  7235 .. 7573  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  8129 .. 9088  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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