Basic Vector Information
Cre-excised pCOIN DV 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 ℃.
Cre-excised pCOIN DV vector Sequence
LOCUS 40924_515 6228 bp DNA circular SYN 17-DEC-2018 DEFINITION Mammalian vector Cre-excised pCOIN DV, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6228) AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 6228) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM REFERENCE 3 (bases 1 to 6228) TITLE Direct Submission REFERENCE 4 (bases 1 to 6228) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6228 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 35..277 /label=chicken beta-globin 5'HS4 insulator core sequence /note="chicken beta-globin 5'HS4 insulator core sequence" misc_feature 278..520 /label=chicken beta-globin 5'HS4 insulator core sequence /note="chicken beta-globin 5'HS4 insulator core sequence" promoter 546..1045 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" regulatory 1059..1068 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" misc_feature 1070..1117 /label=FRT mutant /note="FRT mutant" primer_bind complement(1162..1178) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1186..1202) /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(1210..1240) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1255..1276) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1564..2152) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2326..3183) /label=AmpR /note="beta-lactamase" promoter complement(3184..3288) /label=AmpR promoter primer_bind 3762..3778 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 3801..3848 /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." misc_feature 3865..3968 /label=Ad SA /note="adenovirus major late transcript splice acceptor (Friedrich and Soriano, 1991)" primer_bind 4001..4017 /label=SK primer /note="common sequencing primer, one of multiple similar variants" protein_bind 4031..4064 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." primer_bind 4139..4155 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 4162..4286 /label=attR4 /note="recombination site for the Gateway(R) LR reaction" CDS complement(4330..4632) /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" CDS complement(4977..5633) /label=CmR /note="chloramphenicol acetyltransferase" promoter complement(5687..5717) /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" protein_bind complement(5743..5866) /label=attR3 /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(5894..5910) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" polyA_signal 5959..6183 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal"
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