Plasmid Files

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PciI  (4664) 1 site

PciI is inhibited by nonionic detergents.

EcoO109I  (3844) 1 site

Sticky ends from different EcoO109I sites may not be compatible.

RsrII  (3262) 1 site

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  (2979) 1 site

Sticky ends from different BsrDI sites may not be compatible.

PflFI  (2864) 1 site

The 1-base overhangs produced by PflFI may be hard to ligate. Sticky ends from different PflFI sites may not be compatible.

Tth111I  (2864) 1 site

The 1-base overhangs produced by Tth111I may be hard to ligate. Sticky ends from different Tth111I sites may not be compatible.

FspI  (2848) 1 site

PluTI  (2749) 1 site

Efficient cleavage requires at least two copies of the PluTI recognition sequence.

SfoI  (2747) 1 site

NarI  (2746) 1 site

Efficient cleavage requires at least two copies of the NarI recognition sequence.

KasI  (2745) 1 site

EagI  (2652) 1 site

BspDI  (2586) 1 site

* Blocked by Dam methylation.

ClaI  (2586) 1 site

* Blocked by Dam methylation.

SfiI  (2521) 1 site

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

NdeI  (234) 1 site

Prolonged incubation with NdeI may lead to removal of additional nucleotides.

SnaBI  (340) 1 site

NheI  (591) 1 site

BmtI  (595) 1 site

AfeI  (596) 1 site

AgeI  (600) 1 site

AgeI quickly loses activity at 37°C, but can be used at 25°C for long incubations.

AleI  (613) 1 site

BsrGI  (653) 1 site

BsrGI is typically used at 37°C, but is even more active at 60°C.

BbsI  (911) 1 site

Sticky ends from different BbsI sites may not be compatible. BbsI gradually loses activity when stored at -20°C.

BsmBI  (1042) 1 site

Sticky ends from different BsmBI sites may not be compatible.

PflMI  (1209) 1 site

Sticky ends from different PflMI sites may not be compatible.

PshAI  (1243) 1 site

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.

BspEI  (1321) 1 site

BglII  (1330) 1 site

PaeR7I  (1334) 1 site

PaeR7I does not recognize the sequence CTCTCGAG.

XhoI  (1334) 1 site

Eco53kI  (1339) 1 site

SacI  (1341) 1 site

HindIII  (1343) 1 site

EcoRI  (1350) 1 site

PstI  (1359) 1 site

SalI  (1360) 1 site

Acc65I  (1366) 1 site

KpnI  (1370) 1 site

SacII  (1373) 1 site

Efficient cleavage requires at least two copies of the SacII recognition sequence.

PspOMI  (1374) 1 site

TspMI  (1377) 1 site

XmaI  (1377) 1 site

Efficient cleavage requires at least two copies of the XmaI recognition sequence. Full cleavage with XmaI may require a long incubation.

ApaI  (1378) 1 site

ApaI can be used between 25°C and 37°C.

SmaI  (1379) 1 site

SmaI can be used at 37°C for brief incubations.

BamHI  (1381) 1 site

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.

XbaI  (1393) 1 site

* Blocked by Dam methylation.

BclI  (1403) 1 site

* Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C.

MfeI  (1496) 1 site

HpaI  (1509) 1 site

BtsαI  (1585) 1 site

Sticky ends from different BtsI sites may not be compatible.

MluI  (1632) 1 site

SexAI  (2335) 1 site

* Blocked by Dcm methylation. Sticky ends from different SexAI sites may not be compatible.

NeoR/KanR 2618 .. 3412  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

NeoR/KanR 2618 .. 3412  =  795 bp 264 amino acids  =  29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®)

FusionRed 613 .. 1308  =  696 bp 232 amino acids  =  25.6 kDa Product: monomeric red fluorescent protein optimized for use as a fusion tag (Shemiakina et al., 2012) mammalian codon-optimized

FusionRed 613 .. 1308  =  696 bp 232 amino acids  =  25.6 kDa Product: monomeric red fluorescent protein optimized for use as a fusion tag (Shemiakina et al., 2012) mammalian codon-optimized

ori 4020 .. 4608  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

ori 4020 .. 4608  =  589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication

f1 ori 1638 .. 2093  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

f1 ori 1638 .. 2093  =  456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis

SV40 promoter 2226 .. 2583  =  358 bp SV40 enhancer and early promoter

SV40 promoter 2226 .. 2583  =  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 1510 .. 1631  =  122 bp SV40 polyadenylation signal

SV40 poly(A) signal 1510 .. 1631  =  122 bp SV40 polyadenylation signal

AmpR promoter 2120 .. 2224  =  105 bp

AmpR promoter 2120 .. 2224  =  105 bp

MCS 1321 .. 1386  =  66 bp multiple cloning site

MCS 1321 .. 1386  =  66 bp multiple cloning site

HSV TK poly(A) signal 3644 .. 3691  =  48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)

HSV TK poly(A) signal 3644 .. 3691  =  48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)

SV40 ori 2434 .. 2569  =  136 bp SV40 origin of replication

SV40 ori 2434 .. 2569  =  136 bp SV40 origin of replication

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