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Plasmid Files

pAmCyan1-C1

Vector for fusing AmCyan1 to the N-terminus of a partner protein.

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pAmCyan1-C1 Sequence and MappAmCyan1-C1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 PciI (4643) ApaLI (4329) BsaI (3714) PfoI (3500) RsrII (3241) BsrDI (2958) PflFI - Tth111I (2843) FspI (2827) PluTI (2728) SfoI (2726) NarI (2725) KasI (2724) EagI (2631) BspDI * - ClaI * (2565) BseRI (2543) SfiI (2500) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) BstEII (753) BbsI (1039) KflI - PpuMI (1047) BmgBI (1098) BstXI - PmlI (1197) EcoNI (1231) BsgI (1270) AleI (1285) BspEI (1300) BglII (1309) PaeR7I - XhoI (1313) Eco53kI (1318) SacI (1320) HindIII (1322) EcoRI (1329) SalI (1339) AccI (1340) Acc65I (1345) KpnI (1349) SacII (1352) PspOMI (1353) TspMI - XmaI (1356) ApaI (1357) SmaI (1358) BamHI (1360) XbaI * (1372) stop codons BclI * (1382) MfeI (1475) HpaI (1488) Bts α I (1564) MluI (1611) SexAI * (2314) pAmCyan1-C1 4701 bp
PciI  (4643)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4329)
1 site
G T G C A C C A C G T G
BsaI  (3714)
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.
PfoI  (3500)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
RsrII  (3241)
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.
BsrDI  (2958)
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.
PflFI  (2843)
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  (2843)
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.
FspI  (2827)
1 site
T G C G C A A C G C G T
PluTI  (2728)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (2726)
1 site
G G C G C C C C G C G G
NarI  (2725)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (2724)
1 site
G G C G C C C C G C G G
EagI  (2631)
1 site
C G G C C G G C C G G C
BspDI  (2565)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2565)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BseRI  (2543)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
SfiI  (2500)
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.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SnaBI  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
AgeI  (600)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BstEII  (753)
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.
BbsI  (1039)
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.
KflI  (1047)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1047)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BmgBI  (1098)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BstXI  (1197)
1 site
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.
PmlI  (1197)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
EcoNI  (1231)
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.
BsgI  (1270)
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).
AleI  (1285)
1 site
C A C N N N N G T G G T G N N N N C A C
BspEI  (1300)
1 site
T C C G G A A G G C C T
BglII  (1309)
1 site
A G A T C T T C T A G A
PaeR7I  (1313)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1313)
1 site
C T C G A G G A G C T C
Eco53kI  (1318)
1 site
G A G C T C C T C G A G
SacI  (1320)
1 site
G A G C T C C T C G A G
HindIII  (1322)
1 site
A A G C T T T T C G A A
EcoRI  (1329)
1 site
G A A T T C C T T A A G
SalI  (1339)
1 site
G T C G A C C A G C T G
AccI  (1340)
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.
Acc65I  (1345)
1 site
G G T A C C C C A T G G
KpnI  (1349)
1 site
G G T A C C C C A T G G
SacII  (1352)
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.
PspOMI  (1353)
1 site
G G G C C C C C C G G G
TspMI  (1356)
1 site
C C C G G G G G G C C C
XmaI  (1356)
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.
ApaI  (1357)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (1358)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (1360)
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.
XbaI  (1372)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1382)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MfeI  (1475)
1 site
C A A T T G G T T A A C
HpaI  (1488)
1 site
G T T A A C C A A T T G
BtsαI  (1564)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
MluI  (1611)
1 site
A C G C G T T G C G C A
SexAI  (2314)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
NeoR/KanR
2597 .. 3391  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2597 .. 3391  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
AmCyan1
613 .. 1299  =  687 bp
229 amino acids  =  25.3 kDa
Product: Anemonia majano cyan fluorescent protein
mammalian codon-optimized
AmCyan1
613 .. 1299  =  687 bp
229 amino acids  =  25.3 kDa
Product: Anemonia majano cyan fluorescent protein
mammalian codon-optimized
ori
3999 .. 4587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3999 .. 4587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1617 .. 2072  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1617 .. 2072  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2205 .. 2562  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2205 .. 2562  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1489 .. 1610  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1489 .. 1610  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2099 .. 2203  =  105 bp
AmpR promoter
2099 .. 2203  =  105 bp
MCS
1300 .. 1365  =  66 bp
multiple cloning site of fluorescent protein plasmids
MCS
1300 .. 1365  =  66 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3623 .. 3670  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3623 .. 3670  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
stop codons
1374 .. 1384  =  11 bp
stop codons in all three reading frames
stop codons
1374 .. 1384  =  11 bp
stop codons in all three reading frames
SV40 ori
2413 .. 2548  =  136 bp
SV40 origin of replication
SV40 ori
2413 .. 2548  =  136 bp
SV40 origin of replication
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