Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | Lentiviral-FOP-dGFP-reporter | Antibiotic Resistance | Ampicillin |
| Length | 6922 bp | Type | Mammalian Expression, Lentiviral |
| Copy Number | High Copy | Cloning Method | Restriction Enzyme |
Lentiviral-FOP-dGFP-reporter 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 ℃.
Lentiviral-FOP-dGFP-reporter vector Sequence
LOCUS Exported 6922 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Lentiviral-FOP-dGFP-reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6922)
AUTHORS Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz
L, Nusse R, Weissman IL
TITLE A role for Wnt signalling in self-renewal of haematopoietic stem
cells.
JOURNAL Nature. 2003 May 22. 423(6938):409-14.
PUBMED 12717450
REFERENCE 2 (bases 1 to 6922)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2003 May 22. 423(6938):409-14."
FEATURES Location/Qualifiers
source 1..6922
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 240..260
/label=T3
/note="T3 promoter, forward primer"
promoter 242..260
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 288..514
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 515..695
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 742..867
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1360..1593
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1778..1822
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
regulatory 2156..2165
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2181..2190
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2187..3032
/codon_start=1
/product="EGFP destabilized by fusion to residues 422-461
of mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours"
/label=d2EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
"
primer_bind complement(2232..2253)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2493..2512)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2840..2861
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3053..3641
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3106..3126)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3524..3535)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3728..3961
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4033..4154
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4070..4089)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4124..4143
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4194..4329
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4256..4275
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4349..4368)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4350..4368)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4378..4395)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4378..4394)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4387..4409)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4536..4991
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4623..4642)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4833..4854
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5017..5121
/gene="bla"
/label=AmpR promoter
CDS 5122..5982
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5340..5359)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6153..6741
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6642..6661
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6895..6912
/label=L4440
/note="L4440 vector, forward primer"
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