| 载体基本信息 |
| 载体名称 |
pFlagCMV2-EFP |
载体抗性 |
Ampicillin |
| 载体长度 |
7087 bp |
载体类型 |
Mammalian Expression |
| 拷贝数 |
High Copy |
启动子 |
CMV |
| 克隆方法 |
Restriction Enzyme |
5引物 |
CMV-F |
| 3引物 |
hGH-pA-R (5-CCAGCTTGGTTCCCAATAGA-3) |
pFlagCMV2-EFP 载体载体图谱
pFlagCMV2-EFP 载体载体序列
LOCUS Exported 7087 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFlagCMV2-EFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7087)
AUTHORS Zou W, Zhang DE
TITLE The interferon-inducible ubiquitin-protein isopeptide ligase (E3)
EFP also functions as an ISG15 E3 ligase.
JOURNAL J Biol Chem. 2006 Feb 17. 281(7):3989-94.
PUBMED 16352599
REFERENCE 2 (bases 1 to 7087)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2006 Feb 17. 281(7):3989-94."
FEATURES Location/Qualifiers
source 1..7087
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 618..641
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 3111..3733
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3310..3329)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3757..3777)
/label=pBABE 3
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3762..4091
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3942..4077
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4004..4023
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4129..4148)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4130..4148)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4162..4178)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4162..4178)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4175..4197)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4186..4202
/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(4210..4240)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4255..4276
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4393..4410)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4564..5152)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4644..4663)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5323..6183)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5946..5965
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6184..6288)
/gene="bla"
/label=AmpR promoter
rep_origin 6315..6770
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6402..6421)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6612..6633
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6896..6918
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6910..6927
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6911..6927
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"