Basic Vector Information
MSCV JMJD3 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 ℃.
MSCV JMJD3 vector Sequence
LOCUS 40924_2069 11521 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11521)
AUTHORS Sen GL, Webster DE, Barragan DI, Chang HY, Khavari PA
TITLE Control of differentiation in a self-renewing mammalian tissue by
the histone demethylase JMJD3.
JOURNAL Genes Dev. 2008 Jul 15. 22(14):1865-70.
PUBMED 18628393
REFERENCE 2 (bases 1 to 11521)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11521)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev.
2008 Jul 15. 22(14):1865-70."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11521
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(63..80)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(996..1853)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1854..1958)
/label=AmpR promoter
primer_bind 2026..2044
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2082..2104)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2204..2223
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2417..2439
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2567..2586
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 2786..3302
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 3366..3707
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 3774..4190
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
regulatory 4203..4212
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4212..4235
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
CDS 4248..4274
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
protein_bind 4290..4314
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(4735..4746)
/codon_start=1
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
/translation="IEGR"
CDS 5095..5121
/codon_start=1
/label=9xHis
/note="9xHis affinity tag"
/translation="HHHHHHHHH"
CDS 6585..6611
/codon_start=1
/label=9xHis
/note="9xHis affinity tag"
/translation="HHHHHHHHH"
protein_bind complement(9408..9432)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 9462..10014
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(9606..9623)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 9833..9852
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 10026..10622
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
LTR 10701..11215
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 11377..11393
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(11401..11431)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(11446..11467)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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