Basic Vector Information
- Vector Name:
- pLV.DsRed
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 9186 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Goncalves MA, de Vries AA, Holkers M, van de Watering MJ, van der Velde I, van Nierop GP, Valerio D, Knaan-Shanzer S.
- Promoter:
- EF-1α
pLV.DsRed 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 ℃.
pLV.DsRed vector Sequence
LOCUS 40924_28886 9186 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pLV.DsRed, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9186)
AUTHORS Goncalves MA, de Vries AA, Holkers M, van de Watering MJ, van der
Velde I, van Nierop GP, Valerio D, Knaan-Shanzer S.
TITLE Human mesenchymal stem cells ectopically expressing full-length
dystrophin can complement Duchenne muscular dystrophy myotubes by
cell fusion
JOURNAL Hum. Mol. Genet. 15 (2), 213-221 (2006)
PUBMED 16321987
REFERENCE 2 (bases 1 to 9186)
AUTHORS Goncalves MA, Swildens J, Holkers M, Narain A, van Nierop GP, van de
Watering MJ, Knaan-Shanzer S, de Vries AA.
TITLE Genetic complementation of human muscle cells via directed stem cell
fusion
JOURNAL Mol. Ther. 16 (4), 741-748 (2008)
PUBMED 18334989
REFERENCE 3 (bases 1 to 9186)
AUTHORS Goncalves MA, de Vries AA.
TITLE Direct Submission
JOURNAL Submitted (23-JUL-2007) Molecular Cell Biology, Leiden University
Medical Center, Einthovenweg 20, Leiden 2333 ZC, The Netherlands
REFERENCE 4 (bases 1 to 9186)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 9186)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Hum. Mol.
Genet."; date: "2006"; volume: "15"; issue: "2"; pages: "213-221"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Mol.
Ther."; date: "2008"; volume: "16"; issue: "4"; pages: "741-748"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(23-JUL-2007) Molecular Cell Biology, Leiden University Medical
Center, Einthovenweg 20, Leiden 2333 ZC, The Netherlands"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9186
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=5' LTR
/note="5' long terminal repeat (LTR) from HIV-1"
misc_feature 681..808
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1459..1692
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1789..1906
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3196..4377
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
LTR 4485..4753
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
promoter complement(4790..4808)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4900..5574
/codon_start=1
/label=DsRed-Express
/note="rapidly maturing tetrameric variant of DsRed
fluorescent protein (Bevis and Glick, 2002)"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
LFL"
repeat_region 5625..5859
/label=HIV-1 3' SIN LTR
/note="HIV-1 3' SIN LTR"
LTR 5626..5858
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5883..5901)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5922..5938)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5946..5962)
/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(5970..6000)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6015..6036)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6324..6912)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7304..7960)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(7961..8063)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
promoter complement(8383..8487)
/label=AmpR promoter
rep_origin complement(8513..8968)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 9110..9126
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 9136..9154
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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