Clover-Geminin(1-110) vector (V011877)

MSDS

Basic Vector Information

Vector Name:: Clover-Geminin(1-110)

Antibiotic Resistance:: Ampicillin

Length:: 7475 bp

Type:: Mammalian Expression, Lentiviral

Replication origin:: ori

Copy Number:: High Copy

Promoter:: U6

5' Primer:: agctggtttagtgaaccgtcagatc

3' Primer:: ggaaccggaacccttaaaca

Clover-Geminin(1-110) vector Vector Map

View Map in NovoBuilder

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V011877                 7475 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V011877
VERSION     V011877
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."
            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"
                     /label="Protein Tat"
     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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