Basic Vector Information
- Vector Name:
- PIR2-m1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3846 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM.
- Promoter:
- Pc
PIR2-m1 vector Vector Map
Plasmid Resuspension 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 ℃.
PIR2-m1 vector Sequence
LOCUS 40924_25641 3846 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PIR2-m1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3846)
AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank
LM.
TITLE A Tn7-based device for calibrated heterologous gene expression in
Pseudomonas putida
JOURNAL ACS Synth Biol (2015) In press
PUBMED 26133359
REFERENCE 2 (bases 1 to 3846)
AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank
LM.
TITLE Direct Submission
JOURNAL Submitted (15-JUN-2015) System Biology, Centro Nacional de
Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE 3 (bases 1 to 3846)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3846)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2015) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-JUN-2015) System Biology, Centro Nacional de Biotecnologia,
C/Darwin 3, Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3846
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 56..112
/label=MCS
/note="pUC18/19 multiple cloning site"
CDS 127..840
/codon_start=1
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Pedelacq et al., 2006)"
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 854..948
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
misc_feature 953..1151
/label=Tn7R transposase recognition site
/note="Tn7R transposase recognition site"
CDS 1280..2032
/codon_start=1
/gene="aphA"
/product="aminoglycoside 3'-phosphotransferase"
/label=aphA
/note="kanamycin resistance marker"
/protein_id="AKQ71012.1"
/translation="MLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVT
DEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALA
VFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHK
LLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSL
QKRLFQKYGIDNPDMNKLQFHLMLDEFF"
gene 1280..2032
/gene="aphA"
/label=aphA
/note="aminoglycoside 3'-phosphotransferase"
oriT 2232..2340
/label=oriT
/note="incP origin of transfer"
rep_origin 2359..2747
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
mobile_element 2759..2924
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
terminator complement(2938..3024)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3093..3623)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(3812..3840)
/label=Pc promoter
/note="class 1 integron promoter"
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