pCold- ProS2

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  • Model: PVT0607
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pCold-ProS2

PVT0607   2ug

 

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

     1021 catttggcgg ggattttttt atttgttttc aggaaataaa taatcgatcg cgtaataaaa

     1081 tctattatta tttttgtgaa gaataaattt gggtgcaatg agaatgcgca ggccctttcg

     1141 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt

     1201 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg

     1261 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt

     1321 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat

     1381 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata

     1441 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt

     1501 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc

     1561 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa

     1621 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg

     1681 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt

     1741 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga

     1801 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgtcag

     1861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt

     1921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa

     1981 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt

     2041 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt

     2101 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt

     2161 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg

     2221 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga

     2281 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa

     2341 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga

     2401 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa

     2461 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca

     2521 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta

     2581 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac

     2641 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc

     2701 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag

     2761 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga

     2821 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt

     2881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata

     2941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag

     3001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa

     3061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt

     3121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc

     3181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa

     3241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa

     3301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc

     3361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa

     3421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa

     3481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg

     3541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc

     3601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg

     3661 ctcacatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg

     3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct

     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac

     3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag

     3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc

     3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa

     4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc

     4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc

     4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca

     4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc

     4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt

     4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag

     4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta

     4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc

     4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt

     4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc

     4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac

     4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc

     4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg

     4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc

     4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt

     4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag

     4981 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgc

//

 

Caution:
Product is for research use only!

 

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