Basic Vector Information
- Vector Name:
- 35SpGW
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 5355 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Siqueira Reis R.
- Promoter:
- CaMV 35S (enhanced)
35SpGW 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 ℃.
35SpGW vector Sequence
LOCUS 62056_116 5355 bp DNA circular SYN 26-AUG-2019
DEFINITION Cloning vector 35SpGW, complete sequence.
ACCESSION MK450604
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5355)
AUTHORS Siqueira Reis R.
TITLE CMV 35S promoter-driven Gateway(R) plasmid for protoplast
transformation
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5355)
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 5355)
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"Unpublished"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..5355
/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 358..457
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
primer_bind 550..566
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 666..1337
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 1397..1521
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1546..1576
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1630..2286
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2631..2933
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2977..3101)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3167..3414
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind complement(3463..3562)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
promoter 3684..3754
/label=AmpR promoter
CDS 3755..4612
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4705..5293
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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