Basic Vector Information
The pCDF1-MCS2-EF1-copGFP Vector contains the full-length copGFP gene with optimized human codons for high level of expression of the fluorescent protein from the CMV promoter in mammalian cells. The copGFP marker is a novel natural green monomeric GFP-like protein from copepod (Pontellina sp.). The copGFP protein is a non-toxic, non-aggregating protein with fast protein maturation, high stability at a wide range of pH (pH 4-12), and does not require any additional cofactors or substrates. The copGFP protein has very bright fluorescence that exceeds at least 1.3 times the brightness of EGFP, the widely used Aequorea victoria GFP mutant. The copGFP protein emits green fluorescence with the following characteristics:emission wavelength max – 502 nm; excitation wavelength max – 482 nm; quantum yield – 0.6; extinction coefficient – 70,000 M-1 cm-1
pCDF1-MCS2-EF1-copGFP 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 ℃.
pCDF1-MCS2-EF1-copGFP vector Sequence
LOCUS 40924_9431 6771 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6771)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6771)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6771
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 133..336
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1479..1682
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1839..2050
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2063..2331
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2387..3052
/codon_start=1
/label=CopGFP
/note="green fluorescent protein 2 from Pontellina plumata,
also known as ppluGFP2 (Shagin et al., 2004)"
/translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
"
misc_feature 3131..3719
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 4055..4189
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 4195..4330
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4368..4384)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4392..4408)
/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)."
promoter complement(4416..4446)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4461..4482)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4770..5358)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5532..6389)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6390..6494)
/label=AmpR promoter
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