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Plasmid Files

pACYC177

Plasmid containing the p15A origin of replication and a kanamycin resistance gene.

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pACYC177 Sequence and MappACYC177.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 AcuI (3820) ApaLI (3814) BciVI (3649) AatII (3567) ZraI (3565) EcoO109I (3506) BbsI (3503) PsiI (3485) DrdI (3403) BamHI (3320) BstEII (3279) PflMI (2617) HindIII (2472) BsmBI (2370) AsiSI (2354) EcoNI (2266) SmaI (2228) TspMI - XmaI (2226) BspDI - ClaI (2045) NruI (2011) HincII (3) TatI (62) ScaI (64) SfcI (299) PstI (303) FspI (322) NmeAIII (398) BglI (427) BpmI (476) BsaI (479) BmrI (505) AhdI (545) BanI (592) BstBI (853) AlwNI (992) NspI (1017) SacII (1342) PfoI * (1382) SgrAI (1502) BsgI (1527) AccI (1578) BstZ17I (1579) NheI (1588) BmtI (1592) DraIII (1823) PaeR7I - PspXI - XhoI (1952) BanII (2009) pACYC177 3941 bp
AcuI  (3820)
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).
ApaLI  (3814)
1 site
G T G C A C C A C G T G
BciVI  (3649)
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.
AatII  (3567)
1 site
G A C G T C C T G C A G
ZraI  (3565)
1 site
G A C G T C C T G C A G
EcoO109I  (3506)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BbsI  (3503)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PsiI  (3485)
1 site
T T A T A A A A T A T T
DrdI  (3403)
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.
BamHI  (3320)
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.
BstEII  (3279)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PflMI  (2617)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
HindIII  (2472)
1 site
A A G C T T T T C G A A
BsmBI  (2370)
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.
AsiSI  (2354)
1 site
G C G A T C G C C G C T A G C G
EcoNI  (2266)
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.
SmaI  (2228)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (2226)
1 site
C C C G G G G G G C C C
XmaI  (2226)
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.
BspDI  (2045)
1 site
A T C G A T T A G C T A
ClaI  (2045)
1 site
A T C G A T T A G C T A
NruI  (2011)
1 site
T C G C G A A G C G C T
HincII  (3)
1 site
G T Y R A C C A R Y T G
TatI  (62)
1 site
W G T A C W W C A T G W
ScaI  (64)
1 site
A G T A C T T C A T G A
SfcI  (299)
1 site
C T R Y A G G A Y R T C

Sticky ends from different SfcI sites may not be compatible.
SfcI quickly loses activity at 37°C, but can be used at 25°C for long
incubations.
PstI  (303)
1 site
C T G C A G G A C G T C
FspI  (322)
1 site
T G C G C A A C G C G T
NmeAIII  (398)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (427)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BpmI  (476)
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.
BsaI  (479)
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.
BmrI  (505)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
AhdI  (545)
1 site
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.
BanI  (592)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
BstBI  (853)
1 site
T T C G A A A A G C T T
AlwNI  (992)
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.
NspI  (1017)
1 site
R C A T G Y Y G T A C R
SacII  (1342)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
PfoI  (1382)
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  (1502)
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.
BsgI  (1527)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AccI  (1578)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BstZ17I  (1579)
1 site
G T A T A C C A T A T G
NheI  (1588)
1 site
G C T A G C C G A T C G
BmtI  (1592)
1 site
G C T A G C C G A T C G
DraIII  (1823)
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.
PaeR7I  (1952)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1952)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1952)
1 site
C T C G A G G A G C T C
BanII  (2009)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
AmpR
3699 .. 618  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3699 .. 3767  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3699 .. 618  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3768 .. 618  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3699 .. 618  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
KanR
1923 .. 2738  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
1923 .. 2738  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
p15A ori
783 .. 1328  =  546 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
p15A ori
783 .. 1328  =  546 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
AmpR promoter
3594 .. 3698  =  105 bp
AmpR promoter
3594 .. 3698  =  105 bp
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