pCold- ProS2 Plasmid

$216.00

  • Model: PVT0607
  • 50 Units in Stock
Ask a question


Add to Cart:

pCold-ProS2

Search name

pCold-ProS2,Plasmid pCold-ProS2,pCold-ProS2 vector

 

pCold-ProS2 Information

Promoter: CSPA promoter
Replicon: ColE1 ori
Plasmid Category: E.coli; pCold Series Plasmids
Plasmid size: 5025bp
Plasmid Plasmid: ProS2
Prokaryotic resistance: Ampicillin Amp
Cloning Strains: E. coli DH5α
Culture conditions: 37°C, aerobic, LB
Expression host: E. coli
Culture conditions: 37°C, aerobic, LB
5' Sequencing Primer: pCold-F: 5-ACGCCATATCGCCGAAAGG-3
3' Sequencing Primer: pCold-R: 5-GGCAGGGATCTTAGATTCTG-3

 

pCold-ProS2 Description

pCold ProS2 DNA is a fused cold shock expression vector that uses Protein S derived from the Gram-negative Myxococcus xanthus as a soluble label. This Protein S is present in the spore coat of myxobacteria and consists of 173 amino acid residues and is a very stable soluble protein. By linking the two N-terminal domains of Protein S in tandem and combining ProS2 Tag, the stability and solubility of the fusion protein can be improved. In addition, since the interaction between the target protein and ProS2-Tag is low, the two can be effectively separated. The pCold ProS2 DNA Vector inserts a 5' non-coding region (5'-UTR), a translational enhancement element (TEE), a His-Tag sequence, a ProS2-Tag, and a multiple cloning site (MCS) downstream of the cspA promoter. The cleavage sites of HRV3C Protease, Thrombin, and Factor Xa are mainly between the ProS2-Tag and the multiple cloning site (MCS) to eliminate the fusion protein tag. In addition, the lac operator is inserted downstream of the cspA promoter, allowing the expression of the gene of interest to be tightly regulated. Since the promoter of this product is of E. coli origin, most E. coli strains can be used as expression hosts. In conclusion, the cold shock technology provided by pCold ProS2 DNAVector can promote the correct folding of proteins and increase the solubility of target proteins. It is a good choice for the expression of some difficult target proteins.

 

pCold-ProS2 Sequence

LOCUS       Exported                5025 bp ds-DNA     circular SYN 19-JAN-2017
DEFINITION  synthetic circular DNA
FEATURES             Location/Qualifiers
     source          1..5025
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    84..100
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             271..288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    289..840
                     /label=ProS2
     CDS             850..873
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             883..900
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             907..918
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(1329..1784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1884..1988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1989..2849
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3020..3608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3780..4862)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4863..4940)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcacat ggcaaatatt
      301 accgttttct ataacgaaga cttccagggt aagcaggtcg atctgccgcc tggcaactat
      361 acccgcgccc agttggcggc gctgggcatc gagaataata ccatcagctc ggtgaaggtg
      421 ccgcctggcg tgaaggctat cctgtaccag aacgatggtt tcgccggcga ccagatcgaa
      481 gtggtggcca atgccgagga gttgggcccg ctgaataata acgtctccag catccgcgtc
      541 atctccgtgc ccgtgcagcc gcgcatggca aatattaccg ttttctataa cgaagacttc
      601 cagggtaagc aggtcgatct gccgcctggc aactataccc gcgcccagtt ggcggcgctg
      661 ggcatcgaga ataataccat cagctcggtg aaggtgccgc ctggcgtgaa ggctatcctc
      721 taccagaacg atggtttcgc cggcgaccag atcgaagtgg tggccaatgc cgaggagctg
      781 ggtccgctga ataataacgt ctccagcatc cgcgtcatct ccgtgccggt gcagccgagg
      841 tccgcgggtc tggaagttct gttccagggg ccctccgcgg gtctggtgcc acgcggtagt
      901 ggtggtatcg aaggtaggca tatggagctc ggtaccctcg agggatccga attcaagctt
      961 gtcgacctgc agtctagata ggtaatctct gcttaaaagc acagaatcta agatccctgc
No customer comments for the moment.

Add A Comment

Related Products

Cart  

No products

Total $0.00

Prices don't include postage.

Cart