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pBV220-anti-CD47 nanobody vector (V014075) Gene synthesis in pBV220-anti-CD47 nanobody backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V014075 pBV220-anti-CD47 nanobody In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Engineered E. coliwith Fe₃O₄ nanoparticles are magnetically manipulated to lyse in tumors via heat-sensitive promoters, releasing an anti-CD47 nanobody. This activates immune responses against primary and distal tumors.

Vector Name:
pBV220-anti-CD47 nanobody
Antibiotic Resistance:
Ampicillin
Length:
4057 bp
Type:
Protein expression, Antibody expression
Replication origin:
ori
Host:
E. coli
Promoter:
PL,PR
Growth Strain(s):
Stbl3
Growth Temperature:
30℃

pBV220-anti-CD47 nanobody vector Map

Copy Sequence View Map
pBV220-anti-CD47 nanobody4057 bp60012001800240030003600anti-CD47 nanobodyrrnB T1 terminatorrrnB T2 terminatorAmpR promoterAmpRorilambda repressor (ts)

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

  • Ma X, Liang X, Li Y, Feng Q, Cheng K, Ma N, Zhu F, Guo X, Yue Y, Liu G, Zhang T, Liang J, Ren L, Zhao X, Nie G. Modular-designed engineered bacteria for precision tumor immunotherapy via spatiotemporal manipulation by magnetic field. Nat Commun. 2023 Mar 23;14(1):1606. doi: 10.1038/s41467-023-37225-1. Erratum in: Nat Commun. 2023 Jul 10;14(1):4067. doi: 10.1038/s41467-023-39906-3. PMID: 36959204; PMCID: PMC10036336.

pBV220-anti-CD47 nanobody vector Sequence

LOCUS       Exported                4057 bp DNA     circular SYN 03-JAN-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4057)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4057)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             64..456
                     /codon_start=1
                     /label=anti-CD47 nanobody
                     /translation="MAEVQLQASGGGLVQAGGSLRLSCAASGFTNERSQNMGWFRQAPG
                     KEREGVACISSRVVECWYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCAAPRA
                     ITGTSFVGEHMCDYWGQGTQVTVSSGR"
     terminator      687..773
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      865..892
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        911..1002
                     /label=AmpR promoter
     CDS             1003..1860
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2034..2622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2977..3687)
                     /codon_start=1
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda 
                     repressor"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"