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Plasmid Files

pSB1C3

High copy BioBrick assembly plasmid with a chloramphenicol resistance marker, for shipping parts to the Registry of Standard Biological Parts.

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pSB1C3 Sequence and MappSB1C3.dna
Map and Sequence File   
Sequence Author:  iGEM
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 XbaI (2064) EcoRI (2049) AatII (1943) ZraI (1941) BsaHI (1940) VF2 primer (1931 .. 1950) BsaAI (1912) MslI (1889) TsoI (1706) PvuII (1705) BspEI (1605) BsrDI (1588) AclI (1516) BpmI (1485) BsmBI (1377) PflMI * (1376) PasI (1370) MscI (1338) BtgI - NcoI - StyI (1300) SspI (1293) EcoRV (1153) BanII - SacI (1149) Eco53kI (1147) SpeI (2) PstI (20) his operon terminator BsrBI (146) VR primer (155 .. 174) AflIII - PciI (213) NspI (217) DrdI (321) BciVI (416) HaeII (461) BseYI (517) PspFI (521) ApaLI (527) AlwNI (629) AcuI (761) pSB1C3 2070 bp
XbaI  (2064)
1 site
T C T A G A A G A T C T
EcoRI  (2049)
1 site
G A A T T C C T T A A G
AatII  (1943)
1 site
G A C G T C C T G C A G
ZraI  (1941)
1 site
G A C G T C C T G C A G
BsaHI  (1940)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
BsaAI  (1912)
1 site
Y A C G T R R T G C A Y
MslI  (1889)
1 site
C A Y N N N N R T G G T R N N N N Y A C
TsoI  (1706)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PvuII  (1705)
1 site
C A G C T G G T C G A C
BspEI  (1605)
1 site
T C C G G A A G G C C T
BsrDI  (1588)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
AclI  (1516)
1 site
A A C G T T T T G C A A
BpmI  (1485)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BsmBI  (1377)
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.
PflMI  (1376)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
PasI  (1370)
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.
MscI  (1338)
1 site
T G G C C A A C C G G T
BtgI  (1300)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1300)
1 site
C C A T G G G G T A C C
StyI  (1300)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
SspI  (1293)
1 site
A A T A T T T T A T A A
EcoRV  (1153)
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.
BanII  (1149)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (1149)
1 site
G A G C T C C T C G A G
Eco53kI  (1147)
1 site
G A G C T C C T C G A G
SpeI  (2)
1 site
A C T A G T T G A T C A
PstI  (20)
1 site
C T G C A G G A C G T C
BsrBI  (146)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures
up to 50°C.
AflIII  (213)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (213)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (217)
1 site
R C A T G Y Y G T A C R
DrdI  (321)
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.
BciVI  (416)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
HaeII  (461)
1 site
R G C G C Y Y C G C G R
BseYI  (517)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
PspFI  (521)
1 site
C C C A G C G G G T C G
ApaLI  (527)
1 site
G T G C A C C A C G T G
AlwNI  (629)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AcuI  (761)
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).
VF2 primer
20-mer  /  50% GC
1 binding site
1931 .. 1950  =  20 annealed bases
Tm  =  56°C
forward primer for sequencing/amplifying BioBrick
parts
VR primer
20-mer  /  50% GC
1 binding site
155 .. 174  =  20 annealed bases
Tm  =  57°C
reverse primer for sequencing/amplifying BioBrick
parts
CmR
1160 .. 1819  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1160 .. 1819  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
274 .. 862  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
274 .. 862  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lambda t0 terminator
1045 .. 1139  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
1045 .. 1139  =  95 bp
transcription terminator from phage lambda
his operon terminator
22 .. 79  =  58 bp
This putative transcriptin terminator from the E. coli
his
operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined.
his operon terminator
22 .. 79  =  58 bp
This putative transcriptin terminator from the E. coli
his
operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined.
bacterial terminator
2003 .. 2046  =  44 bp
putative bacterial transcription terminator
bacterial terminator
2003 .. 2046  =  44 bp
putative bacterial transcription terminator
BioBrick prefix
2049 .. 2070  =  22 bp
BioBrick prefix
2049 .. 2070  =  22 bp
BioBrick suffix
1 .. 21  =  21 bp
BioBrick suffix
1 .. 21  =  21 bp
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