Basic Vector Information
- Vector Name:
- pTSL
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4255 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Taylor NM, Prokhorov NS, Guerrero-Ferreira RC, Shneider MM, Browning C, Goldie KN, Stahlberg H, Leiman PG.
pTSL 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 ℃.
pTSL vector Sequence
LOCUS 40924_44394 4255 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTSL, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4255)
AUTHORS Taylor NM, Prokhorov NS, Guerrero-Ferreira RC, Shneider MM, Browning
C, Goldie KN, Stahlberg H, Leiman PG.
TITLE Structure of the T4 baseplate and its function in triggering sheath
contraction
JOURNAL Nature 533 (7603), 346-352 (2016)
PUBMED 27193680
REFERENCE 2 (bases 1 to 4255)
AUTHORS Shneider MM, Miroshikov KA.
TITLE Direct Submission
JOURNAL Submitted (17-DEC-2015) Laboratory of Molecular Bioengineering,
IBCH, Miklukho-Maklaya str. 16/10, Moscow 117997, Russian Federation
REFERENCE 3 (bases 1 to 4255)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4255)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature";
date: "2016"; volume: "533"; issue: "7603"; pages: "346-352"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-DEC-2015) Laboratory of Molecular Bioengineering, IBCH,
Miklukho-Maklaya str. 16/10, Moscow 117997, Russian Federation"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4255
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS complement(207..227)
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS complement(786..803)
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
RBS complement(828..850)
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind complement(865..889)
/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)."
promoter complement(890..908)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1417..1605
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 1710..1852
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(2038..2626)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2800..3657)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3658..3762)
/label=AmpR promoter
rep_origin complement(3789..4244)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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