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
| 载体基本信息 | |||
| 载体名称 | pCDF1-MCS2-EF1-copGFP | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6771 bp | 载体类型 | Lentiviral vectors |
| 启动子 | CMV | 克隆方法 | Enzyme digestion and ligation |
| 5引物 | LNCX: AGCTCGTTTAGTGAACCGTCAGATC | 3引物 | EF1a-R: TTTCGCCCTAACTTCGTGAT |
| 载体标签 | copGFP | ||
pCDF1-MCS2-EF1-copGFP 载体载体图谱
pCDF1-MCS2-EF1-copGFP 载体载体序列
LOCUS Exported 6771 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6771)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6771
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 133..336
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1479..1682
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1839..2050
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2063..2331
/note="5 LTR (truncated)"
/note="truncated 5 long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2390..3052
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/note="CopGFP"
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
misc_feature 3131..3719
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(3602..3613)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
polyA_signal 4055..4176
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4195..4330
/note="SV40 ori"
/note="SV40 origin of replication"
primer_bind complement(4368..4384)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4392..4408
/bound_moiety="lac repressor encoded by lacI"
/note="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)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4461..4482
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4770..5358)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5529..6389)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6390..6494)
/gene="bla"
/note="AmpR promoter"
