The pBudCE4.1 vector is designed for the independent expression of two genes from a single plasmid in mammalian cells. Using pBudCE4.1 to generate stable mammalian expression cell lines ensures that there is an equivalent copy number of each gene in the cell. This can eliminate variable expression due to differences in gene copy number. pBudCE4.1 provides expression cassettes withthe following features:
| 载体基本信息 | |||
| 载体名称 | pBudCE4.1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4595 bp | 载体类型 | Mammalian Expression Vectors (Dual Promoter) |
| 筛选标记 | Zeocin | 启动子 | EF-1α, CMV |
| 克隆方法 | Enzyme digestion and ligation | 5引物 | CMVPro-F: 5d[TAATACGACTCACTATAGGG]3 EF1aPro-F:5d[TCAAGCCTCAGACAGTGGTTC]3 |
| 载体标签 | 6xHis, c-Myc, V5 | ||
pBudCE4.1 载体载体图谱
pBudCE4.1 载体载体序列
LOCUS Exported 4595 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 4595)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4595
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 10..389
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 638..656
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 719..748
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 764..781
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 812..933
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(1063..1437)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1456..1503)
/note="EM7 promoter"
/note="synthetic bacterial promoter "
promoter complement(1551..1867)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 1565..1700
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 1889..3051
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2120..3042
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
CDS 3127..3168
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 3178..3195
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 3224..3448
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(3580..4168)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
