Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004477 | pMRE104_PnptII_eCFP | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pMRE104_PnptII_eCFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13484 bp
- Type:
- Tn7 delivery vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Remus-Emsermann MNP., Gisler P, Drissner D.
- Promoter:
- araBAD
pMRE104_PnptII_eCFP 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.
pMRE104_PnptII_eCFP vector Sequence
LOCUS V004477 13484 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004477
VERSION V004477
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13484)
AUTHORS Remus-Emsermann MNP., Gisler P, Drissner D.
TITLE MiniTn7-transposon delivery vectors for inducible or constitutive
fluorescent protein expression in Enterobacteriaceae
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 13484)
AUTHORS Remus-Emsermann MNP., Drissner D.
TITLE Direct Submission
JOURNAL Submitted (23-MAR-2016) Institute for Food Sciences, Agroscope,
Schloss 1, Waedenswil, Zurich 8820, Switzerland
REFERENCE 3 (bases 1 to 13484)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13484)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-MAR-2016) Institute for Food Sciences, Agroscope, Schloss 1,
Waedenswil, Zurich 8820, Switzerland"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13484
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(149..1024)
/label="araC"
/note="L-arabinose regulatory protein"
promoter 1051..1335
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 1453..2271
/gene="tnsA"
/label="Transposon Tn7 transposition protein TnsA"
/note="Transposon Tn7 transposition protein TnsA from
Escherichia coli. Accession#: P13988"
CDS 2261..4366
/gene="tnsB"
/label="Transposon Tn7 transposition protein TnsB"
/note="Transposon Tn7 transposition protein TnsB from
Escherichia coli. Accession#: P13989"
CDS 4366..6030
/gene="tnsC"
/label="Transposon Tn7 transposition protein TnsC"
/note="Transposon Tn7 transposition protein TnsC from
Escherichia coli. Accession#: P05846"
CDS 6036..7562
/codon_start=1
/gene="tnsD"
/product="TnsD"
/label="tnsD"
/note="Tn7 transposition site-selector"
/protein_id="AMZ00002.1"
/translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
gene 6036..7562
/gene="tnsD"
/label="tnsD"
terminator 8196..8282
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 8374..8401
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 8420..8511
/label="AmpR promoter"
CDS 8512..9369
/label="AmpR"
/note="beta-lactamase"
oriT complement(9543..9652)
/direction=LEFT
/label="oriT"
/note="incP origin of transfer"
rep_origin 10392..10614
/label="pSC101 ori"
/note="low-copy replication origin that requires the Rep101
protein"
CDS 10662..11609
/label="Rep101(Ts)"
/note="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
mobile_element 12045..12210
/label="Tn7L"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
CDS 12509..13225
/label="ECFP"
/note="enhanced CFP"
repeat_region 13273..13484
/label="Tn7 right end"
/note="Tn7 right end"