Basic Vector Information
p35S-loxP-Zeo 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 ℃.
p35S-loxP-Zeo vector Sequence
LOCUS 40924_2632 5167 bp DNA circular SYN 18-DEC-2018 DEFINITION Moss transformation vector p35S-loxP-Zeo DNA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5167) AUTHORS Aoyama T, Hiwatashi Y, Hasebe M. TITLE Moss transformation vector p35S-loxP-Zeo JOURNAL Unpublished REFERENCE 2 (bases 1 to 5167) AUTHORS Aoyama T, Hiwatashi Y, Hasebe M. TITLE Direct Submission JOURNAL Submitted (07-JAN-2010) Contact:Yuji Hiwatashi National Institute for Basic Biology, Evolutionary Biology; Myodaiji, Nishigonaka, 38, Okazaki, Aichi 444-8585, Japan REFERENCE 3 (bases 1 to 5167) TITLE Direct Submission REFERENCE 4 (bases 1 to 5167) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (07-JAN-2010) Contact:Yuji Hiwatashi National Institute for Basic Biology, Evolutionary Biology; Myodaiji, Nishigonaka, 38, Okazaki, Aichi 444-8585, Japan" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5167 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(3..458) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 670..686 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 708..741 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_recomb 708..714 /label=loxP /note="loxP" primer_bind 816..832 /label=SK primer /note="common sequencing primer, one of multiple similar variants" polyA_signal complement(1344..1520) /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(1593..1964) /codon_start=1 /label=BleoR /note="antibiotic-binding protein" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVLVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" promoter complement(1971..2313) /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" protein_bind complement(2854..2887) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." promoter complement(2979..2997) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3018..3034) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3042..3058) /label=lac operator /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(3066..3096) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3111..3132) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3420..4008) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4182..5039) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5040..5144) /label=AmpR promoter
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