Basic Vector Information
- Vector Name:
- pTVDll42AEmerald
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6572 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Blanco R, Geudens I, Stanchi F, Mathivet T, Jones M, Bentley K, Gerhardt H.
- Promoter:
- mPGK
pTVDll42AEmerald 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 ℃.
pTVDll42AEmerald vector Sequence
LOCUS 40924_44519 6572 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTVDll42AEmerald, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6572)
AUTHORS Blanco R, Geudens I, Stanchi F, Mathivet T, Jones M, Bentley K,
Gerhardt H.
TITLE Collective endothelial Dll4/Notch dynamics switch between vessel
branching and expansion
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6572)
AUTHORS Ubezio B.
TITLE Direct Submission
JOURNAL Submitted (25-JUN-2013) Vascular Biology Laboratory, Cancer Research
UK, 44 Lincoln's Inn Fields, London, London WC2A 3LY, United Kingdom
REFERENCE 3 (bases 1 to 6572)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6572)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-JUN-2013) Vascular Biology Laboratory, Cancer Research UK, 44
Lincoln's Inn Fields, London, London WC2A 3LY, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6572
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 471..575
/label=AmpR promoter
CDS 576..1433
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1607..2195
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2483..2504
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2519..2549
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2557..2573
/label=lac operator
/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)."
primer_bind 2581..2597
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2618..2636
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 3215..3928
/codon_start=1
/label=EmGFP
/note="Emerald GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELY"
protein_bind 4006..4039
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 4104..4603
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
promoter 4615..4662
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 4681..5481
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 5522..5746
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter complement(6379..6397)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6404..6420)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(join(6561..6572,1..444))
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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