pRetroX-IRES-ZsGreen1 is a bicistronic, fl uorescent, retroviral vector that allows both a gene of interest and the ZsGreen1 gene to be translated from a single bicistronic mRNA. pRetroXIRES-ZsGreen1 is designed for effi cient delivery and selection (by fl ow cytometry or other methods) of stably transduced mammalian cells expressing the ZsGreen1 fl uorescent protein and the protein of interest. This vector can be used to obtain stable cell lines without timeconsuming drug and clonal selection. ZsGreen1 is derived from the reef coral Zoanthus sp. green fl uorescent protein (ZsGreen) and is easily detected with standard FITC fi lter sets.
| 载体基本信息 | |||
| 载体名称 | pRetroX-IRES-ZsGreen1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5953 bp | 载体类型 | Viral Expression & Packaging Vectors |
| 载体来源 | Clontech | 拷贝数 | High copy number |
| 启动子 | CMV | ||
pRetroX-IRES-ZsGreen1 载体载体图谱
pRetroX-IRES-ZsGreen1 载体载体序列
LOCUS Exported 5953 bp ds-DNA circular SYN 29-9-2014
DEFINITION Bicistronic retroviral vector for expressing a gene together with
the fluorescent protein ZsGreen1.
ACCESSION .
VERSION .
KEYWORDS pRetroX-IRES-ZsGreen1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5953)
AUTHORS Clontech
TITLE Direct Submission
COMMENT
FEATURES Location/Qualifiers
source 1..5953
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
LTR 1..592
/note="5 LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 653..658
/note="splice donor"
/note="splice donor site (beginning of intron)"
misc_feature 655..1012
/note="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
intron 1075..1278
/note="EF-1-alpha intron A"
/note="truncated intron upstream of the start codon of
human EF-1-alpha"
exon 1279..1308
/note="EF-1-alpha exon"
/note="truncated exon 2 from human EF-1-alpha, beginning
with a GT-to-CC mutation that reduces splicing efficiency"
misc_feature 1362..1389
/note="MCS"
/note="multiple cloning site"
misc_feature 1393..1965
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 1967..2662
/codon_start=1
/product="Zoanthus green fluorescent protein"
/note="ZsGreen1"
/note="mammalian codon-optimized"
/translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
AIASGSALP"
LTR 2716..3309
/note="3 LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
rep_origin complement(3727..4315)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4486..5346)
/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(5347..5451)
/gene="bla"
/note="AmpR promoter"
