Basic Vector Information
- Vector Name:
- pNL-DT5R
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12076 bp
- Type:
- HIV-1 vector
- Replication origin:
- ori
- Source/Author:
- Kamada K, Igarashi T, Martin MA, Khamsri B, Hatcho K, Yamashita T, Fujita M, Uchiyama T, Adachi A.
pNL-DT5R 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 ℃.
pNL-DT5R vector Sequence
LOCUS 40924_33332 12076 bp DNA circular SYN 18-DEC-2018
DEFINITION HIV-1 vector pNL-DT5R DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12076)
AUTHORS Kamada K, Igarashi T, Martin MA, Khamsri B, Hatcho K, Yamashita T,
Fujita M, Uchiyama T, Adachi A.
TITLE Generation of HIV-1 derivatives that productively infect macaque
monkey lymphoid cells
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 103 (45), 16959-16964 (2006)
PUBMED 17065315
REFERENCE 2 (bases 1 to 12076)
AUTHORS Kamada K, Adachi A.
TITLE Direct Submission
JOURNAL Submitted (20-JUL-2006) Contact:Kazuya Kamada Kobe university school
of medicine, Department of microbiology; 7-5-1, Kusunokicho,
Chuo-ku, Kobe, Hyogo 650-0017, Japan
REFERENCE 3 (bases 1 to 12076)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12076)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A."; date: "2006"; volume: "103"; issue: "45"; pages:
"16959-16964"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-JUL-2006) Contact:Kazuya Kamada Kobe university school of
medicine, Department of microbiology; 7-5-1, Kusunokicho, Chuo-ku,
Kobe, Hyogo 650-0017, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12076
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 377..634
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
CDS 790..2286
/codon_start=1
/product="gag polyprotein"
/label=gag polyprotein
/note="Cyclophilin binding loop (PVHAGPIAP) in capsid
region was replaced to SIVmac analogues (PQPAPQQ)"
/protein_id="BAF34641.1"
/translation="MGARASVLSGGELDKWEKIRLRPGGKKQYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTIAVLYCVHQRIDVKDTKEALDKI
EEEQNKSKKKAQQAAADTGNNSQVSQNYPIVQNLQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRLHPQPAPQ
QGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTSIL
DIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPGAT
LEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEGHI
AKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAPPE
ESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 2079..5087
/label=HIV-1 pol
/note="pol protein from human immunodeficiency virus 1"
misc_feature 5092..5096
/note="artificially inserted sequence for generating Sma I
site"
CDS 5097..5741
/codon_start=1
/product="vif protein"
/label=vif protein
/note="derived from SIVmac239"
/protein_id="BAF34643.1"
/translation="MEEEKRWIAVPTWRIPERLERWHSLIKYLKYKTKDLQKVCYVPHF
KVGWAWWTCSRVIFPLQEGSHLEVQGYWHLTPEKGWLSTYAVRITWYSKNFWTDVTPNY
ADILLHSTYFPCFTAGEVRRAIRGEQLLSCCRFPRAHKYQVPSLQYLALKVVSDVRSQG
ENPTWKQWRRDNRRGLRMAKQNSRGDKQRGGKPPTKGANFPGLAKVLGILA"
CDS 5744..6031
/gene="vpr"
/label=vpr
/note="Protein Vpr from Human immunodeficiency virus type 1
group M subtype B (isolate NY5). Accession#: P12520"
CDS 6015..6188
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate BH5). Accession#: P04612"
CDS 6406..8964
/gene="env"
/label=env
/note="Envelope glycoprotein gp160 from Human
immunodeficiency virus type 1 group M subtype B (isolate
MFA). Accession#: P19551"
CDS 8972..8998
/gene="nef"
/label=nef
/note="Protein Nef from Human immunodeficiency virus type 1
group M subtype D (isolate Z84). Accession#: P12481"
LTR 9261..9894
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(9919..9935)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9943..9959
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(9967..9997)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(10012..10033)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(10321..10909)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11083..11940)
/label=AmpR
/note="beta-lactamase"
promoter complement(11941..12045)
/label=AmpR promoter
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