Basic Vector Information
- Vector Name:
- GWpNF
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 8687 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Siqueira Reis R.
- Promoter:
- lac UV5
GWpNF 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 ℃.
GWpNF vector Sequence
LOCUS 62056_1116 8687 bp DNA circular SYN 26-AUG-2019
DEFINITION Cloning vector GWpNF, complete sequence.
ACCESSION MK450605
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8687)
AUTHORS Siqueira Reis R.
TITLE Gateway(R) sites followed by nano and firefly (nLuc/Fluc) dual
luciferase for protoplast transformation
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8687)
AUTHORS Siqueira Reis R.
TITLE Direct Submission
JOURNAL Submitted (26-JAN-2019) Department of Plant Molecular Biology,
University of Lausanne, Quartier UNIL-Sorge, Batiment Biophore,
Lausanne, Vaud 1015, Switzerland
REFERENCE 3 (bases 1 to 8687)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-JAN-2019) Department of Plant Molecular Biology, University of
Lausanne, Quartier UNIL-Sorge, Batiment Biophore, Lausanne, Vaud
1015, Switzerland"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8687
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 103..189
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 368..492
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 517..547
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 601..1257
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 1602..1904
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(1948..2072)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2086..2151
/codon_start=1
/label=F2A
/note="2A peptide from foot-and-mouth disease virus
polyprotein"
/translation="VKQLLNFDLLKLAGDVESNPGP"
CDS 2170..2682
/codon_start=1
/label=Nluc
/note="NanoLuc(R) luciferase"
/translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
RLCERILA"
CDS 2683..2709
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
terminator 2732..2980
/label=HSP terminator
/note="efficient transcription terminator from the
Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya
et al., 2010)"
protein_bind complement(2989..3013)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
CDS 4748..6397
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
terminator 6427..6679
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 7016..7086
/label=AmpR promoter
CDS 7087..7944
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 8037..8625
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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