Basic Vector Information
STOP-eGFP-ROSA26TV 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 ℃.
STOP-eGFP-ROSA26TV vector Sequence
LOCUS STOP-eGFP-ROSA26 14367 bp DNA circular SYN 11-SEP-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS STOP-eGFP-ROSA26TV.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14367)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 14367)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14367
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 5..1086
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1093..1196
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
protein_bind 1207..1240
/label=Cre recombinase binding site
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal complement(1247..1295)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1305..2105)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
intron 2919..2984
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 3114..3134
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 3559..3693
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(3713..3746)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind 3779..3826
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 3848..4427
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4428..5144
/label=EGFP
/note="enhanced GFP"
protein_bind 5172..5219
/label=FLP recombinase from the Saccharomyces
cerevisi
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(5244..5260)
/label=SK primer
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 5279..5503
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 5596..6379
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
promoter 9897..10387
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 10414..10983
/label=DTA
/note="diphteria toxin Fragment A"
intron 11060..11125
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 11199..11423
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter complement(11522..11540)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11561..11577)
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 11585..11601
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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(11609..11639)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(11654..11675)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11963..12551)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12725..13582)
/label=AmpR
/note="beta-lactamase"
promoter complement(13583..13687)
/label=AmpR promoter
rep_origin complement(13713..14168)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 14310..14326
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 14336..14354
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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