The primer for sequencing out the 5 end of the GFP based constructs (GFP/EGFP, CFP, YFP) is 5- GTCTTGTAGTTGCCGTCGTC -3
| 载体基本信息 | |||
| 载体名称 | pcDNA3-YFP | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6159 bp | 载体类型 | Mammalian Expression Vectors |
| 筛选标记 | G418, neo | 拷贝数 | High copy number |
| 启动子 | CMV | 克隆方法 | Enzyme digestion and ligation |
| 5引物 | T7 Fwd:5d[TAATACGACTCACTATAGGG]3 | 载体标签 | His Tag (6x), c-Myc Epitope Tag |
pcDNA3-YFP 载体载体图谱
pcDNA3-YFP 载体载体序列
LOCUS Exported 6159 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 6159)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6159
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 977..1696
/codon_start=1
/product="enhanced YFP"
/note="EYFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
promoter complement(1712..1730)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 1756..1980
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 2026..2454
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2468..2797
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2648..2783
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2864..3658
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3832..3953
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(4002..4018)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4026..4042
/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(4050..4080)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4095..4116
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4404..4992)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5163..6023)
/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(6024..6128)
/gene="bla"
/note="AmpR promoter"
