Basic Vector Information
ATG-less TV 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 ℃.
ATG-less TV vector Sequence
LOCUS 40924_305 6741 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector ATG-less TV, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6741)
AUTHORS Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y,
Konishi H.
TITLE A system for the measurement of gene targeting efficiency in human
cell lines using an antibiotic resistance-GFP fusion gene
JOURNAL BioTechniques 53 (3), 141-152 (2012)
PUBMED 22963476
REFERENCE 2 (bases 1 to 6741)
AUTHORS Konishi H.
TITLE Direct Submission
JOURNAL Submitted (07-JAN-2012) Aichi Medical University School of Medicine,
1-1 Yazako-Karimta, Nagakute, Aichi 480-1195, Japan
REFERENCE 3 (bases 1 to 6741)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6741)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"BioTechniques"; date: "2012"; volume: "53"; issue: "3"; pages:
"141-152"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(07-JAN-2012) Aichi Medical University School of Medicine, 1-1
Yazako-Karimta, Nagakute, Aichi 480-1195, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6741
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
CDS 180..896
/label=EGFP
/note="enhanced GFP"
primer_bind 900..916
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 918..951
/label=loxP
/note="loxP"
protein_bind 918..951
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
intron 989..1218
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
misc_feature 1259..1809
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 1823..2614
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal complement(2639..2773)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(2870..2903)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(2956..2972)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2975..3993
/note="partial SV40 largeT; an internal portion of SV40
largeT exon 2"
repeat_region 4004..4144
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
rep_origin 4219..4674
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4956..5060
/label=AmpR promoter
CDS 5061..5918
/label=AmpR
/note="beta-lactamase"
rep_origin 6092..6680
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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