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Plasmid Files

pVLT33

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

 
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pVLT33.dna
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BamHI (9702) KpnI (9697) Acc65I (9693) SacI (9691) Eco53kI (9689) EcoRI (9681) MauBI (9406) AclI (9289) MluI (8949) ApaI (8746) PspOMI (8742) HpaI (8447) BsmBI (8334) FspAI (7670) EcoNI (7520) SgrAI (5432) PasI (4953) XbaI (9708) SbfI (9724) DraIII (414) BglII (764) EagI (834) DrdI (955) PflFI - Tth111I (1046) RsrII (1444) BstBI (1610) NruI (1783) Bsu36I (1804) SfiI (2572) BstZ17I (3947) pVLT33 9762 bp
BamHI  (9702)
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.
KpnI  (9697)
1 site
G G T A C C C C A T G G
Acc65I  (9693)
1 site
G G T A C C C C A T G G
SacI  (9691)
1 site
G A G C T C C T C G A G
Eco53kI  (9689)
1 site
G A G C T C C T C G A G
EcoRI  (9681)
1 site
G A A T T C C T T A A G
MauBI  (9406)
1 site
C G C G C G C G G C G C G C G C
AclI  (9289)
1 site
A A C G T T T T G C A A
MluI  (8949)
1 site
A C G C G T T G C G C A
ApaI  (8746)
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  (8742)
1 site
G G G C C C C C C G G G
HpaI  (8447)
1 site
G T T A A C C A A T T G
BsmBI  (8334)
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.
FspAI  (7670)
1 site
R T G C G C A Y Y A C G C G T R
EcoNI  (7520)
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.
SgrAI  (5432)
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.
PasI  (4953)
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.
XbaI  (9708)
1 site
T C T A G A A G A T C T
SbfI  (9724)
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.
BglII  (764)
1 site
A G A T C T T C T A G A
EagI  (834)
1 site
C G G C C G G C C G G C
DrdI  (955)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PflFI  (1046)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1046)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
RsrII  (1444)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (1610)
1 site
T T C G A A A A G C T T
NruI  (1783)
1 site
T C G C G A A G C G C T
Bsu36I  (1804)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SfiI  (2572)
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  (3947)
1 site
G T A T A C C A T A T G
lacI
8222 .. 9304  =  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
8222 .. 9304  =  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
4786 .. 5757  =  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
4786 .. 5757  =  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
7094 .. 7945  =  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
7094 .. 7945  =  852 bp
283 amino acids  =  31.1 kDa
Product: replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
NeoR/KanR
800 .. 1594  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
800 .. 1594  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
RSF1010 oriV
2724 .. 3118  =  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
2724 .. 3118  =  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
2090 .. 2205  =  116 bp
T4 transcription termination signals
T4 terminators
2090 .. 2205  =  116 bp
T4 transcription termination signals
RSF1010 oriT
3460 .. 3547  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
RSF1010 oriT
3460 .. 3547  =  88 bp
origin of transfer of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
lacIq promoter
9305 .. 9382  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
9305 .. 9382  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
9681 .. 9737  =  57 bp
pUC18/19 multiple cloning site
MCS
9681 .. 9737  =  57 bp
pUC18/19 multiple cloning site
tac promoter
9612 .. 9640  =  29 bp
   Segment 1:  -35  
   9612 .. 9617  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9612 .. 9640  =  29 bp
   Segment 2:  
   9618 .. 9633  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9612 .. 9640  =  29 bp
   Segment 3:  -10  
   9634 .. 9640  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
9612 .. 9640  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
lac operator
9648 .. 9664  =  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
9648 .. 9664  =  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
6268 .. 7107  =  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
6268 .. 7107  =  840 bp
279 amino acids  =  29.9 kDa
Product: replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)
ORF:  1615 .. 2037  =  423 bp
ORF:  140 amino acids  =  15.5 kDa
ORF:  3628 .. 5757  =  2130 bp
ORF:  709 amino acids  =  78.0 kDa
ORF:  6268 .. 7107  =  840 bp
ORF:  279 amino acids  =  29.9 kDa
ORF:  800 .. 1594  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2924 .. 3352  =  429 bp
ORF:  142 amino acids  =  15.3 kDa
ORF:  3470 .. 3877  =  408 bp
ORF:  135 amino acids  =  14.3 kDa
ORF:  4376 .. 4789  =  414 bp
ORF:  137 amino acids  =  15.1 kDa
ORF:  5213 .. 5857  =  645 bp
ORF:  214 amino acids  =  22.8 kDa
ORF:  6995 .. 7945  =  951 bp
ORF:  316 amino acids  =  34.4 kDa
ORF:  972 .. 1358  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2310 .. 2717  =  408 bp
ORF:  135 amino acids  =  15.0 kDa
ORF:  1645 .. 1890  =  246 bp
ORF:  81 amino acids  =  8.4 kDa
ORF:  3145 .. 3429  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  3499 .. 4155  =  657 bp
ORF:  218 amino acids  =  22.9 kDa
ORF:  4348 .. 4686  =  339 bp
ORF:  112 amino acids  =  12.0 kDa
ORF:  4903 .. 5496  =  594 bp
ORF:  197 amino acids  =  20.5 kDa
ORF:  6265 .. 7032  =  768 bp
ORF:  255 amino acids  =  27.0 kDa
ORF:  7435 .. 7776  =  342 bp
ORF:  113 amino acids  =  12.6 kDa
ORF:  2346 .. 2591  =  246 bp
ORF:  81 amino acids  =  9.4 kDa
ORF:  3795 .. 4025  =  231 bp
ORF:  76 amino acids  =  8.4 kDa
ORF:  4692 .. 5045  =  354 bp
ORF:  117 amino acids  =  12.6 kDa
ORF:  6519 .. 6884  =  366 bp
ORF:  121 amino acids  =  13.3 kDa
ORF:  8202 .. 8465  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  9054 .. 9371  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1109 .. 1645  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  2354 .. 2584  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  7328 .. 7666  =  339 bp
ORF:  112 amino acids  =  12.1 kDa
ORF:  8222 .. 9181  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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