Basic Vector Information
pUC.Donor.GFP.dATG 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 ℃.
pUC.Donor.GFP.dATG vector Sequence
LOCUS 40924_44943 4180 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC.Donor.GFP.dATG, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4180)
AUTHORS Holkers M, de Vries AA, Goncalves MA.
TITLE Nonspaced inverted DNA repeats are preferential targets for
homology-directed gene repair in mammalian cells
JOURNAL Nucleic Acids Res. 40 (5), 1984-1999 (2012)
PUBMED 22080552
REFERENCE 2 (bases 1 to 4180)
AUTHORS Holkers M, de Vries AF, Goncalves MAF.V.
TITLE Direct Submission
JOURNAL Submitted (22-MAR-2011) Molecular Cell Biology, Leiden University
Medical Center, Einthovenweg 20, Leiden, Zuid-Holland 2333 ZC, The
Netherlands
REFERENCE 3 (bases 1 to 4180)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4180)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2012"; volume: "40"; issue: "5"; pages:
"1984-1999"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-MAR-2011) Molecular Cell Biology, Leiden University Medical
Center, Einthovenweg 20, Leiden, Zuid-Holland 2333 ZC, The
Netherlands"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4180
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 148..203
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
CDS 567..1250
/codon_start=1
/label=hrGFP
/note="humanized Renilla green fluorescent protein"
/translation="VEEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGNQLVQIRVTKGA
PLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYEDGGLVEIRSDI
NLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVGQVILVYRLNSG
KFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAIAQLTSLGKPLG
SLHEWV"
polyA_signal complement(1285..1406)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1521..1537)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2011..2115
/label=AmpR promoter
CDS 2116..2973
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3147..3735
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4023..4044
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4059..4089
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4097..4113
/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)."
primer_bind 4121..4137
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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