Basic Vector Information
- Vector Name:
- pMBL6attR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3967 bp
- Type:
- Moss gene targeting vector
- Replication origin:
- ori
- Source/Author:
- Kamisugi Y, Cuming AC, Cove DJ.
pMBL6attR 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 ℃.
pMBL6attR vector Sequence
LOCUS 40924_29896 3967 bp DNA circular SYN 18-DEC-2018
DEFINITION Moss gene targeting vector pMBL6attR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3967)
AUTHORS Kamisugi Y, Cuming AC, Cove DJ.
TITLE Parameters determining the efficiency of gene targeting in the moss
Physcomitrella patens
JOURNAL Nucleic Acids Res. 33 (19), E173 (2005)
PUBMED 16282584
REFERENCE 2 (bases 1 to 3967)
AUTHORS Kamisugi Y, Cove DJ, Cuming AC.
TITLE Direct Submission
JOURNAL Submitted (01-OCT-2005) Centre for Plant Sciences, Leeds University,
Leeds, West Yorkshire LS2 9JT, UK
REFERENCE 3 (bases 1 to 3967)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3967)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2005"; volume: "33"; issue: "19"; pages: "E173"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-OCT-2005) Centre for Plant Sciences, Leeds University, Leeds,
West Yorkshire LS2 9JT, UK"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3967
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 394..696
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(740..864)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(923..939)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(947..963)
/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(971..1001)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1016..1037)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1325..1913)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2087..2944)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2945..3049)
/label=AmpR promoter
protein_bind 3127..3251
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 3276..3306
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS join(3360..3967,1..49)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
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