Basic Vector Information
Clover-Geminin(1-110) 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 ℃.
Clover-Geminin(1-110) vector Sequence
LOCUS Clover-Geminin(1 7475 bp DNA circular SYN 13-MAY-2021
DEFINITION Fluorescent probe for M/G1 transition.
ACCESSION .
VERSION .
KEYWORDS Clover-Geminin(1-110).
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7475)
AUTHORS Bajar BT, Lam AJ, Badiee RK, Oh YH, Chu J, Zhou XX, Kim N, Kim BB,
Chung M, Yablonovitch AL, Cruz BF, Kulalert K, Tao JJ, Meyer T, Su
XD, Lin MZ
TITLE Fluorescent indicators for simultaneous reporting of all four cell
cycle phases.
JOURNAL Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045.
PUBMED 27798610
REFERENCE 2 (bases 1 to 7475)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7475)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7475
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 63..366
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 368..566
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 584..764
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 811..936
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1435..1668
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1853..1897
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 2046..2087
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2195..2312
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2371..2684
/label=U6 promoter
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
primer_bind 2701..2717
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2759..2792
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
enhancer 2908..3211
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3212..3415
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3447..4163
/label=Clover
/note="bright green-yellow fluorescent protein derived from
GFP (Lam et al., 2012)"
CDS 4194..4523
/codon_start=1
/product="degron consisting of residues 1-110 of human
Geminin (Zielke and Edgar, 2015)"
/label=Gem1 (1-110)
/translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
KRRKAL"
protein_bind complement(4562..4595)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 4651..5239
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5242..5258)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 5481..5661
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5723..6311)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6485..7342)
/label=AmpR
/note="beta-lactamase"
promoter complement(7343..7447)
/label=AmpR promoter
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