Product Details

Purity

>85% (SDS-PAGE)

Target Names

seqA

Uniprot No.

P0AFY8

Alternative Names

seqA; b0687; JW0674; Negative modulator of initiation of replication

Species

Escherichia coli (strain K12)

Expression Region

1-181

Target Protein Sequence

MKTIEVDDEL YSYIASHTKH IGESASDILR RMLKFSAASQ PAAPVTKEVR VASPAIVEAK PVKTIKDKVR AMRELLLSDE YAEQKRAVNR FMLLLSTLYS LDAQAFAEAT ESLHGRTRVY FAADEQTLLK NGNQTKPKHV PGTPYWVITN TNTGRKCSMI EHIMQSMQFP AELIEKVCGT I

Protein Length

full length protein

Tag Info

Tag type will be determined during the manufacturing process.

The tag type will be determined during production process. If you have specified tag type, please tell us and we will develop the specified tag preferentially.

Form

Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer before Lyophilization

Tris/PBS-based buffer, 6% Trehalose, pH 8.0

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Note: All of our proteins are default shipped with normal blue ice packs, if you request to ship with dry ice, please communicate with us in advance and extra fees will be charged.

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet

Please contact us to get it.

Target Background

Function

Negative regulator of replication initiation, which contributes to regulation of DNA replication and ensures that replication initiation occurs exactly once per chromosome per cell cycle. Binds to pairs of hemimethylated GATC sequences in the oriC region, thus preventing assembly of replication proteins and re-initiation at newly replicated origins. Repression is relieved when the region becomes fully methylated. Can also bind to hemimethylated GATC sequences outside of oriC region. Binds, with less affinity, to fully methylated GATC sites and affects timing of replication. May play a role in chromosome organization and gene regulation.

Gene References into Functions

Data indicate that the SeqA protein is localized at midcell for most of the D-period, but colocalizes with the origin region before cell division and initiation of replication. PMID: 25722374
Data suggest that, in the absence of SeqA, the sister-chromatid cohesion 'safety spacer' is destabilized and completely disappears if the replication fork is inhibited. PMID: 24806348
We propose that SeqA binding results in loose inter-duplex junctions that are resistant to Topo IV cleavage PMID: 23990792
Cell cycle analysis showed that loss of SeqA from the old replication forks did not occur at initiation of the new forks, but instead occurs at a time point coinciding with the end of SeqA-dependent origin sequestration. PMID: 22373925
The number of SeqA foci is not directly governed by the number of replication forks, and supports the idea that new DNA may be 'captured' by existing SeqA structures. PMID: 19371375
well-defined SeqA structures are not found in situations of extensive replication fork repair PMID: 19898675
SeqA protein bound to fully methylated, covalently closed or nicked circular DNA generates positive supercoils in vitro in the presence of topoisomerase I or ligase respectively. PMID: 15458410
These results indicate that SeqA binds hemimethylated nascent DNA segments according to the proceeding of replication forks in the chromosome PMID: 15612935
Data report the crystal structure of the oligomerization domain of SeqA (SeqA-N). PMID: 15933720
Overproduction of SeqA4 led to partially restored initiation synchrony, indicating that origin sequestration may not depend on efficient higher-order multimerization into foci, but do require a high local concentration of SeqA. PMID: 16236133
These results suggest that for survival of replication fork damage, SeqA's repression of replication initiation is more important than its effects on nucleoid organization. PMID: 16556234
it has been demonstrated that two GATC sequences, located 82 and 105 bp downstream of the bacteriophage lambda p(R) transcription start site, are necessary for transcription stimulation by SeqA both in vivo and in vitro PMID: 17005979
investigation of how resetting of oriC to the ORC-like complex is coordinated with SeqA-mediated sequestration; during the sequestration period, SeqA repressed pre-RC assembly while ensuring resetting of E. coli ORC PMID: 17114060
These results indicate that unlike oriC, SeqA modulates lambda DNA replication indirectly, most probably by influencing the stability of the lambda replication complex and the transcriptional activation of ori lambda. PMID: 17464080
An examination of the molecular model of Escherichia coli SeqA protein, a negative regulator of chromosome DNA replication which prevents the overinitiation of replication. PMID: 18849124
seqA mutants have increased LPS phosphorylation and exhibit high levels of chromosomal fragmentation. PMID: 19432803

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Subcellular Location

Cytoplasm. Note=Localization is cell cycle-dependent. Localizes at midcell in newborn cells, then migrates in opposite directions toward cell quarter sites and remains tethered there until the cell divides.

Protein Families

SeqA family

Database Links

KEGG: ecj:JW0674

STRING: 316385.ECDH10B_0752

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Recombinant Escherichia coli Negative modulator of initiation of replication (seqA)

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