Basic Vector Information
- Vector Name:
- pENTR5'_ubi:loxP-EGFP-loxP
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7496 bp
- Type:
- Multisite Gateway 5' entry vector
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- Ubi
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- M13 forward
- 3' Primer:
- M13 reverse
pENTR5'_ubi:loxP-EGFP-loxP 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 ℃.
pENTR5'_ubi:loxP-EGFP-loxP vector Sequence
LOCUS 40924_17537 7496 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7496) AUTHORS Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI TITLE Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. JOURNAL Development. 2011 Jan . 138(1):169-77. PUBMED 21138979 REFERENCE 2 (bases 1 to 7496) TITLE Direct Submission REFERENCE 3 (bases 1 to 7496) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Development. 2011 Jan . 138(1):169-77." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7496 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator complement(268..295) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(387..473) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 592..691 /label=attL1 /note="recombination site for the Gateway(R) LR reaction" promoter 715..4197 /label=Ubi /note="Zebrafish ubiquitin (ubi) promoter" protein_bind 4210..4243 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 4256..4972 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 5098..5219 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5363..5384) /label=F1ori-F /note="F1 origin, forward primer" protein_bind complement(5473..5506) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 5537..5661 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter complement(5738..5756) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5761..5777) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 5890..6696 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 6846..7434 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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