Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V012094 | pJV53 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pJV53
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8801 bp
- Type:
- Bacterial Expression
- Replication origin:
- ori
- Selection Marker:
- Kanamycin
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pJV53 vector Map
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.
pJV53 vector Sequence
LOCUS Exported 8801 bp DNA circular SYN 22-JAN-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8801)
AUTHORS van Kessel JC, Hatfull GF
TITLE Recombineering in Mycobacterium tuberculosis.
JOURNAL Nat Methods. 2007 Feb . 4(2):147-52.
PUBMED 17179933
REFERENCE 2 (bases 1 to 8801)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8801)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8801)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2007 Feb . 4(2):147-52."
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8801
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 1..1884
/mol_type="other DNA"
/clone="pJV53-Cpf1"
/note="derived from Francisella tularensis subsp.
novicida;
codon-optimized FnCpf1 inducing expression vector"
/db_xref="taxon:32630"
/organism="synthetic construct"
source 1885..8801
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 3110..3538
/mol_type="other DNA"
/label=Escherichia coli/Mycobacterium shuttle plasmid
/note="Escherichia coli/Mycobacterium shuttle plasmid"
/db_xref="taxon:930172"
/organism="Shuttle vector pMycoFos"
source 3620..3904
/mol_type="other DNA"
/label=Escherichia coli/Mycobacterium shuttle plasmid
/note="Escherichia coli/Mycobacterium shuttle plasmid"
/db_xref="taxon:930172"
/organism="Shuttle vector pMycoFos"
source 3909..4550
/mol_type="other DNA"
/label=Escherichia coli/Mycobacterium shuttle plasmid
/note="Escherichia coli/Mycobacterium shuttle plasmid"
/db_xref="taxon:930172"
/organism="Shuttle vector pMycoFos"
rep_origin 1..1884
/label=oriM
/note="oriM"
CDS complement(1934..3007)
/codon_start=1
/label=amiC
/translation="MQDGEVEFRVGLVIPLQGPAGIFAPSCEAVAELAAKEVNDRGGLQ
GRKVTIEVLDGGAPGDDVARTVADRLRGHGLDAVTGWHISAVRNRISPVVRDRIPYVYT
SLYEGGERTPGVFCTGETPQIQIAPALAWLRDHFGIRSWCLVGDDYIWPRRSAAAARAY
CRDLDLELRREIYVPYGTDDFRAPVRKAIASGAQAVLMLLVGQDAVLFNREFARAGGHD
RMARFSPLMEENMLLASGAGSTENLYVAAAYFSSLATAGAMDLMGSYVARYGADAPPLN
AMAESCYEGLLALEAIFQRAHSPEIPDLMASAHDVGFDGPRGPMCMRDSQFDQQVYIAS
ADGYDFDILDTLATLDA"
promoter 3045..4568
/label=Acetamidase Promoter
gene 3110..3538
/gene="amiA"
/label=amiA
CDS 3110..3538
/codon_start=1
/transl_table=11
/gene="amiA"
/product="AmiA"
/label=amiA
/note="acetamidase regulator"
/protein_id="ADR02800.1"
/translation="MAAGQQRRPNLLLPLVRLTHLAESAIERVLADSSLKIEDWRVLDE
LAGRRTVPMSDLAQATLITGPTLTRTVDRLVSQGIIYRTADLHDRRRVLVALTPRGRTL
RNRLVDAVAEAECAAFESCGLDVDQLRELVDTTSNLTS"
promoter 3558..3603
/label=P2 promoter
gene 3620..3904
/gene="amiD"
/label=amiD
CDS 3620..3904
/codon_start=1
/transl_table=11
/gene="amiD"
/product="AmiD"
/label=amiD
/note="acetamidase regulator"
/protein_id="ADR02801.1"
/translation="MPTYTFRCSHCGPFDLTCAISERDAAATCPECRTPARRVFGSVGL
TTFTAGHHRAFDAASASAESPTVVKSIPAGADRPRAPRRNPGLPSLPRY"
gene 3909..4550
/gene="amiS"
/label=amiS
CDS 3909..4550
/codon_start=1
/transl_table=11
/gene="amiS"
/product="AmiS"
/label=amiS
/note="acetamidase transporter"
/protein_id="ADR02802.1"
/translation="MGGVGLFYVGAVLIIDGLMLLGRISPRGATPLNFFVGGLQVVTPT
VLILQSGGDAAVIFAASGLYLFGFTYLWVAINNVTDWDGEGLGWFSLFVAIAALGYSWH
AFTAEADPAFGVIWLLWAVLWFMLFLLLGLGHDALGPAVGFVAVAEGVITAAVPAFLIV
SGNWETGPLPAAVIAVIGFAAVVLAYPIGRRLAAPSVTNPPPAALAATTR"
CDS 4639..5583
/codon_start=1
/label=che9c 60
/note="exonuclease VIII from Mycobacterium phage Che9c"
/translation="MSVPTQDGMHRFVDEDVYHADRGSLSVSGAKLLLPPSCPAKFRWE
MDNTRKPKKVWDFGHVAHKLVLGKGAEFEILDPEVHGLKADGTPSEKPTATGMWRKAEA
EARKQGKVPIHVDLFTKAYDMAEKVRQHPTAGPIFANPDGEAEVALYYTDPETGVRLRG
RIDWLTDDIDDYKTSMTANPAELKTKFYKLGYFMQAAWYIDLLVALGLAENPRFRFITQ
EKEPPYVVTPIQYDDEAIEEGRRRNRQAIRLYADCMESGKWPDYSDDVVTISLPSWGLP
RPQTVGDVVTDSYIYDTDPLEEADPIEGDYIYG"
CDS 5576..6637
/codon_start=1
/label=che9c 61
/note="recombinase RecT from Mycobacterium tuberculosis"
/translation="MAENAVTKQDSPKAPETISQVLQVLVPQLARAVPKGMDPDRIARI
VQTEIRKSRNAKAAGIAKQSLDDCTQESFAGALLTSAALGLEPGVNGECYLVPYRDTRR
GVVECQLIIGYQGIVKLFWQHPRASRIDAQWVGANDEFHYTMGLNPTLKHVKAKGDRGN
PVYFYAIVEVTGAEPLWDVFTADEIRELRRGKVGSSGDIKDPQRWMERKTALKQVLKLA
PKTTRLDAAIRADDRPGTDLSQSQALALPSTVKPTADYIDGEIAEPHEVDTPPKSSRAQ
RAQRATAPAPDVQMANPDQLKRLGEIQKAEKYNDADWFKFLADSAGVKATRAADLTFDE
AKAVIDMFDGPNA"
terminator 6702..6748
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS 6909..7721
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 8055..8643
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"