pSMART BAC v2.0

BAC vector with inducible copy number, suitable for cloning toxic inserts.

Sequence Author: Lucigen

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BstXI (7644) BamHI (7632) HindIII (7625) EcoRI (7618) PmlI (7614) AhdI (7606) NotI (7594) SP6 promoter SL1 (7546 .. 7566) BstXI (8) BamHI (12) HindIII (19) EcoRI (26) PmlI (33) AhdI (41) NotI (48) SR4 (84 .. 104) pSMART® BAC v2.0 7648 bp
BstXI  (7644)
2 sites
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BamHI  (7632)
2 sites
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.
HindIII  (7625)
2 sites
A A G C T T T T C G A A
EcoRI  (7618)
2 sites
G A A T T C C T T A A G
PmlI  (7614)
2 sites
C A C G T G G T G C A C
AhdI  (7606)
2 sites
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
NotI  (7594)
2 sites
G C G G C C G C C G C C G G C G
BstXI  (8)
2 sites
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BamHI  (12)
2 sites
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.
HindIII  (19)
2 sites
A A G C T T T T C G A A
EcoRI  (26)
2 sites
G A A T T C C T T A A G
PmlI  (33)
2 sites
C A C G T G G T G C A C
AhdI  (41)
2 sites
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
NotI  (48)
2 sites
G C G G C C G C C G C C G G C G
SL1
21-mer  /  57% GC
1 binding site
7546 .. 7566  =  21 annealed bases
Tm  =  59°C
SR4
21-mer  /  33% GC
1 binding site
84 .. 104  =  21 annealed bases
Tm  =  52°C
sopA
3211 .. 4386  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
sopA
3211 .. 4386  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
repE
4965 .. 5720  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
repE
4965 .. 5720  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
CmR
254 .. 913  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
254 .. 913  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
oriV
6106 .. 6720  =  615 bp
origin of replication for the bacterial F plasmid
oriV
6106 .. 6720  =  615 bp
origin of replication for the bacterial F plasmid
sopC
1694 .. 2167  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
sopC
1694 .. 2167  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
cos
1035 .. 1433  =  399 bp
lambda cos site; allows packaging into phage lambda particles
cos
1035 .. 1433  =  399 bp
lambda cos site; allows packaging into phage lambda particles
incC
4712 .. 4962  =  251 bp
incompatibility region of the bacterial F plasmid
incC
4712 .. 4962  =  251 bp
incompatibility region of the bacterial F plasmid
ori2
5811 .. 6030  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
ori2
5811 .. 6030  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
cat promoter
151 .. 253  =  103 bp
promoter of the E. coli cat gene
cat promoter
151 .. 253  =  103 bp
promoter of the E. coli cat gene
rrnBT1 terminator
102 .. 147  =  46 bp
transcription terminator T1 from the E. coli rrnB gene
rrnBT1 terminator
102 .. 147  =  46 bp
transcription terminator T1 from the E. coli rrnB gene
loxP
984 .. 1017  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
984 .. 1017  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
tonB terminator
927 .. 958  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
927 .. 958  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
T7Te terminator
7487 .. 7514  =  28 bp
phage T7 early transcription terminator
T7Te terminator
7487 .. 7514  =  28 bp
phage T7 early transcription terminator
T7 promoter
61 .. 79  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
61 .. 79  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
7573 .. 7591  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
7573 .. 7591  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
sopB
2240 .. 3211  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
sopB
2240 .. 3211  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
ORF:  254 .. 913  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  7238 .. 7582  =  345 bp
ORF:  114 amino acids  =  13.0 kDa
ORF:  2934 .. 3170  =  237 bp
ORF:  78 amino acids  =  8.3 kDa
ORF:  5331 .. 5564  =  234 bp
ORF:  77 amino acids  =  9.1 kDa
ORF:  2240 .. 3211  =  972 bp
ORF:  323 amino acids  =  35.4 kDa
ORF:  5171 .. 5416  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  5501 .. 5731  =  231 bp
ORF:  76 amino acids  =  8.7 kDa
ORF:  7115 .. 7462  =  348 bp
ORF:  115 amino acids  =  12.9 kDa
ORF:  2461 .. 2859  =  399 bp
ORF:  132 amino acids  =  15.6 kDa
ORF:  3211 .. 4386  =  1176 bp
ORF:  391 amino acids  =  44.1 kDa
ORF:  4965 .. 5720  =  756 bp
ORF:  251 amino acids  =  29.4 kDa
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