Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V003726 | pRK415 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pRK415
- Antibiotic Resistance:
- Tetracycline
- Length:
- 10692 bp
- Type:
- Broad host range expression vector
- Replication origin:
- oriV
- Source/Author:
- Hulter N, Wackernagel W.
- Promoter:
- lac
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pRK415 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.
pRK415 vector Sequence
LOCUS Exported 10692 bp DNA circular SYN 08-SEP-2025
DEFINITION Broad host range expression vector pRK415, complete sequence.
ACCESSION EF437940
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10692)
AUTHORS Hulter N, Wackernagel W.
TITLE Double illegitimate recombination events integrate DNA segments
through two different mechanisms during natural transformation of
Acinetobacter baylyi
JOURNAL Mol. Microbiol. 67 (5), 984-995 (2008)
PUBMED 18194157
REFERENCE 2 (bases 1 to 10692)
AUTHORS Huelter N, Wackernagel W.
TITLE Direct Submission
JOURNAL Submitted (13-FEB-2007) Genetics, Department of Biology and
Environmental Sciences, Carl von Ossietzky University Oldenburg,
Carl-von-Ossietzkystrasse 9-11, Oldenburg 26111, Germany
REFERENCE 3 (bases 1 to 10692)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Microbiol."; date: "2008"; volume: "67"; issue: "5"; pages:
"984-995"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-FEB-2007) Genetics, Department of Biology and Environmental
Sciences, Carl von Ossietzky University Oldenburg,
Carl-von-Ossietzkystrasse 9-11, Oldenburg 26111, Germany"
FEATURES Location/Qualifiers
source 1..10692
/mol_type="other DNA"
/note="derived from plasmid pRK404 GenBank Accession Number
AY204475"
/db_xref="taxon:429723"
/organism="Broad host range expression vector pRK415"
rep_origin 1..712
/label=oriV
/note="incP origin of replication"
rep_origin 276..987
/note="oriV; IncP"
CDS complement(971..1312)
/codon_start=1
/gene="lacZ-alpha"
/product="beta-galactosidase alpha peptide"
/label=lacZ-alpha
/protein_id="ABO32178.1"
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRHASRREKGGQVSGKRQGRNRRAHEGA
SRGKRLVSL"
gene complement(971..1312)
/gene="lacZ-alpha"
/label=lacZ-alpha
primer_bind 1222..1238
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1239..1295
/label=multiple cloning site
/note="multiple cloning site"
primer_bind complement(1308..1324)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1332..1348
/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)."
regulatory 1352..1421
/label=lac promoter
/note="lac promoter"
/regulatory_class="promoter"
promoter complement(1356..1386)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1401..1422
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
gene complement(1654..2279)
/gene="traJ"
/label=traJ
CDS complement(1654..2025)
/codon_start=1
/gene="traJ"
/product="TraJ"
/label=traJ
/note="oriT-recognizing protein"
/protein_id="ABO32179.1"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 2058..2167
/label=incP
/note="incP"
misc_feature 2299..2532
/label=similar to NC_001371 Plasmid ColE1
/note="similar to NC_001371 Plasmid ColE1"
misc_feature 2524..2676
/label=similar to NC_001371 Plasmid ColE1
/note="similar to NC_001371 Plasmid ColE1"
CDS complement(2848..3498)
/codon_start=1
/gene="tetR"
/product="TetR"
/label=tetR
/note="tetracycline resistance regulation"
/protein_id="ABO32180.1"
/translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
gene complement(2848..3498)
/gene="tetR"
/label=tetR
CDS 3604..4803
/codon_start=1
/gene="tetA"
/product="tetracycline resistance protein"
/label=tetA
/protein_id="ABO32181.1"
/translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
"
gene 3604..4803
/gene="tetA"
/label=tetA
CDS complement(4835..5911)
/codon_start=1
/label=upf16.5
/translation="MIKAQIRRLGAADVQDFREIRLSALKKAPEMFGSVYEHEEKKPME
AFAERLRDAVAFGAYIDGEIIGLSVFKQEDGPKDAHKAHLSGVFVEPEQRGRGVAGMLL
RALPAGLLLVMIVRQIPTGIWWMRIFILGALNISLFWSLLFISVYRLPGGVAATVGAVQ
PLMVVFISAALLGSPIRLMAVLGAICGTAGVALLVLTPNAALDPVGVAAGLAGAVSMAF
GTVLTRKWQPPVPLLTFTAWQLAAGGLLLVPVALVFDPPIPMPTGTNVLGLAWLGLIGA
GLTYFLWFRGISRLEPTVVSLLGFLSPGTAVLLGWLFLDQTLSALQIIGVLLVIGSIWL
GQRSNRTPRARIACRKSP"
gene complement(5434..6101)
/gene="upf16.5"
/label=upf16.5
CDS complement(6076..7224)
/codon_start=1
/gene="trfA2"
/product="trans-acting replication A2"
/label=trfA2
/protein_id="ABO32183.1"
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
gene complement(6076..7224)
/gene="trfA2"
/label=trfA2
CDS complement(6076..6933)
/codon_start=1
/gene="trfA1"
/product="trans-acting replication A1"
/label=trfA1
/protein_id="ABO32184.1"
/translation="MTTKKRKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAP
WPDTMRAMPNDTARSALFTTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELV
WQQVLEYAKRTPIGEPITFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRV
GHLESVSLLHRFRVLDRGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRM
FDYFSSHREPYPLKLETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLV
HCKR"
gene complement(6076..6933)
/gene="trfA1"
/label=trfA1
CDS complement(7273..7623)
/codon_start=1
/gene="ssb"
/product="single-stranded DNA binding protein"
/label=ssb
/protein_id="ABO32185.1"
/translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
LDTKAPGGSNQE"
gene complement(7273..7623)
/gene="ssb"
/label=ssb
CDS 7744..8109
/codon_start=1
/gene="trbA"
/product="TrbA"
/label=trbA
/note="regulation of tra gene expression"
/protein_id="ABO32186.1"
/translation="MYNQIFFTNILRLLDERGMTKHELSERAGVSISFLSDLTNGKANP
SLKVMEAIADALETPLPLLLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAF
IVKKWGDDTRKKLRGRL"
gene 7744..8109
/gene="trbA"
/label=trbA
CDS complement(9147..10241)
/codon_start=1
/gene="incC1"
/product="IncC1"
/label=incC1
/note="IncC partitioning protein, long form"
/protein_id="ABO32187.1"
/translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
AARKASKEVRALADYVFTKMEISQ"
gene complement(9147..10241)
/gene="incC1"
/label=incC1
CDS complement(9147..9926)
/codon_start=1
/gene="incC2"
/product="IncC2"
/label=incC2
/note="IncC partitioning protein, short form"
/protein_id="ABO32188.1"
/translation="MKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNAS
YTLKDFATGLHASKLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDA
RELFGANIKALANQGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMV
TTIANVRQKNAKLQFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADA
LASGVPVWKIKKTAARKASKEVRALADYVFTKMEISQ"
gene complement(9147..9926)
/gene="incC2"
/label=incC2
CDS complement(9923..10228)
/codon_start=1
/gene="korA"
/product="KorA"
/label=korA
/note="regulation of gene expression"
/protein_id="ABO32189.1"
/translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
gene complement(9923..10228)
/gene="korA"
/label=korA