Basic Vector Information
- Vector Name:
- TKVBL-GVB
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7480 bp
- Type:
- Retrofitting vector
- Replication origin:
- oriV
- Source/Author:
- Kondo S, Booker M, Perrimon N.
- Promoter:
- copia
TKVBL-GVB 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 ℃.
TKVBL-GVB vector Sequence
LOCUS 40924_48937 7480 bp DNA circular SYN 18-DEC-2018
DEFINITION Retrofitting vector TKVBL-GVB, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7480)
AUTHORS Kondo S, Booker M, Perrimon N.
TITLE Cross-species RNAi rescue platform in Drosophila melanogaster
JOURNAL Genetics 183 (3), 1165-1173 (2009)
PUBMED 19720858
REFERENCE 2 (bases 1 to 7480)
AUTHORS Kondo S, Booker M, Perrimon N.
TITLE Direct Submission
JOURNAL Submitted (19-FEB-2009) Department of Genetics, Harvard Medical
School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA
REFERENCE 3 (bases 1 to 7480)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7480)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genetics";
date: "2009"; volume: "183"; issue: "3"; pages: "1165-1173"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-FEB-2009) Department of Genetics, Harvard Medical School, 77
Avenue Louis Pasteur, Boston, MA 02115, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7480
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1107..1140
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind 1457..1886
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
polyA_signal complement(1894..2028)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(2360..2755)
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
promoter complement(2772..3051)
/label=copia promoter
/note="strong promoter from the Drosophila transposable
element copia (Sinclair et al., 1986)"
polyA_signal complement(3171..3252)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(3381..4097)
/codon_start=1
/label=Venus
/note="yellow fluorescent protein (YFP) with fast and
efficient maturation (Nagai et al., 2002)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
protein_bind 4635..5019
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
CDS complement(5391..6203)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 6327..6962
/label=oriV
/note="incP origin of replication"
protein_bind 6997..7066
/label=attB
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
mobile_element complement(7292..7457)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
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