Basic Vector Information
- Vector Name:
- pLgw V5-EcoDam
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8629 bp
- Type:
- Mammalian Expression, Lentiviral ; DamID
- Replication origin:
- ori
- Selection Marker:
- Zeo marker is outside the LTRs and will not be packaged into virus.
- Promoter:
- hsp70
- 5' Primer:
- pCasper-hs
pLgw V5-EcoDam 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 ℃.
pLgw V5-EcoDam vector Sequence
LOCUS pLgw_V5-EcoDam. 8629 bp DNA circular SYN 13-MAY-2021
DEFINITION Mammalian DamID control lentiviral vector for expression of V5-Dam
only .
ACCESSION .
VERSION .
KEYWORDS pLgw V5-EcoDam.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8629)
AUTHORS Vogel MJ, Guelen L, de Wit E, Peric-Hupkes D, Loden M, Talhout W,
Feenstra M, Abbas B, Classen AK, van Steensel B
TITLE Human heterochromatin proteins form large domains containing
KRAB-ZNF genes.
JOURNAL Genome Res. 2006 Dec;16(12):1493-504. Epub 2006 Oct 12.
PUBMED 17038565
REFERENCE 2 (bases 1 to 8629)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8629)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genome
Res."; date: "2006-12"; volume: "16(12)"; pages: "1493-504. Epub
2006 Oct 12"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8629
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 36..171
/label=SV40 ori
/note="SV40 origin of replication"
promoter 233..280
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 299..670
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 803..936
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(973..989)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(997..1013)
/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(1021..1051)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1066..1087)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1204..1221)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1375..1963)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2137..2994)
/label=AmpR
/note="beta-lactamase"
promoter complement(2995..3099)
/label=AmpR promoter
primer_bind complement(3174..3193)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3365..3744
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3745..3947
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 3962..4142
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4186..4311
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4804..5037
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5222..5266
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 5415..5456
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
promoter 5728..5966
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
protein_bind 6019..6043
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(6119..6143)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
CDS 6154..6195
/label=V5 tag
/note="epitope tag from simian virus 5"
CDS 6214..7047
/gene="dam"
/label=dam
/note="DNA adenine methylase from Escherichia coli (strain
K12). Accession#: P0AEE8"
LTR 7560..7740
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 7772..7996
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8042..8470
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8484..8629
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
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