| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pRetroQ-DsRed-Monomer-N1 |
| 质粒类型: | 逆病毒载体；荧光报告载体 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | CMV IE |
| 载体大小: | 7571 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | DsRed-Monomer(C-端） |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 嘌呤霉素（Puromycin） |
| 克隆菌株: | DH5α |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等），原代细胞 |
| 备注: | pRetroQ-DsRed-Monomer-N1载体是自我失活型逆病毒载体，表达C端DsRed-Monomer融合蛋白； pRetroQ-DsRed-Monomer-N1载体能够高效递送目的基因到靶细胞中，尤其适用于原代细胞和其它难转染的细胞系； DsRed-Monomer是DsRed的单体突变体，与DsRed相比，编码序列经过了人工优化，适用于在哺乳动物细胞中的高水平表达。 |
| 产品目录号: | ZY632507 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 逆转录病毒 |
- ¥3500
- ZYbscience
- 中国/美国
- ZY632507
- 2025年07月12日
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- 详细信息
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pRetroQ-DsRed-Monomer-N1
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pRetroQ-DsRed Monomer-N1载体描述
pRetroQ-DsRed Monomer-N1 is a high-titer, self-inactivating retroviral vector that facilitates efficient delivery and expression of DsRed-monomer (DsRed.M1) as well as N terminal fusions of DsRed monomer to target cells. DsRed.M1 is a monomeric mutant derived from the tetrameric Discosoma sp. red fluorescent protein DsRed (1). DsRed-Monomer contains forty-five amino acid substitutions. When DsRed-Monomer is expressed in mammalian cell cultures, red fluorescent cells can be detected by either fluorescence microscopy or flow cytometry 12–16 hours after transfection or 24–48 hours after infection (DsRed-Monomer excitation and emission maxima = 557 nm and 592 nm, respectively. The DsRed-Monomer coding sequence is human codon-optimized for high expression in mammalian cells (2) The MCS in pRetroQ-DsRed Monomer-N1 lies between the immediate early promoter of CMV (PCMV IE) and the DsRed-Monomer coding sequences. Genes cloned into the MCS are expressed as fusions to the N-terminus of DsRed Monomer when they are in the same reading frame as DsRed Monomer and there are no intervening stop codons. The RetroQ retroviral vector backbone incorporates several unique features. This vector contains a puromycin resistance cassette (Puror) driven by the PGK promoter for selection of positively-infected cells (2).The hybrid 5’ long terminal repeat (LTR) consists of the CMV type I enhancer and the murine sarcoma virus (MSV) promoter. This vector demonstrates high levels of transcription in HEK 293-based packaging cell lines due, in part, to the presence of adenoviral E1A (3–6) in these cells. The self-inactivating feature of the vector is provided by a deletion in the 3’ LTR enhancer region (U3). During reverse transcription of the retroviral RNA, the inactivated 3’ LTR is copied and replaces the 5’ LTR CMV enhancer sequences. This can reduce the phenomenon known as promoter interference (7) and allow more efficient expression.
Additionally, the viral genomic transcript contains the necessary viral RNA processing elements, including the LTRs, packaging signal (ψ+), and tRNA primer-binding site. pRetroQDsRed Monomer-N1 contains a bacterial origin of replication, an E. coli Ampr gene for propagation and selection in bacteria, and an SV40 origin for replication in mammalian cells expressing the SV40 T antigen.
pRetroQ-DsRed Monomer-N1 is designed to efficiently deliver and express fusions to the N terminus of DsRed-Monomer into primary cells or cells that are difficult to transfect. Fusions to the N terminus of DsRed-Monomer retain the fluorescent properties of the native protein, allowing the in vivo localization of the fusion protein. The target gene should be cloned into pDsRed Monomer-N1 so that it is in frame with the DsRed-Monomer coding sequences, with no intervening in-frame stop codons. The inserted gene should include the initiating ATG codon. The recombinant DsRed Monomer vector can be infected or transfected into mammalian cells. If required, stable transformants can be selected using puromycin. pRetroQ DsRed Monomer-N1 can also be used simply to express DsRed Monomer in a cell line of interest (e.g., as an infection marker). Once pRetroQ-DsRed Monomer-N1 is transfected into a packaging cell line (such as the RetroPack PT67 Cell line, AmphoPack-293, EcoPack2-293, Pantropic Expression System , or Retro-X Universal Packaging System, RNA from the vector is packaged into non-infectious, replication-incompetent retroviral particles, since pRetroQ-DsRed Monomer-N1 lacks the structural genes (gag, pol, and env) necessary for particle formation and replication. These genes, however, are stably integrated as part of the packaging cell genome. Once a high-titer supernatant is produced, these retroviral particles can infect target cells and transmit the gene of interest but cannot replicate within these cells due to the absence of viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chances of producing replicationcompetent virus due to recombination events during cell proliferation.
Propagation in E. coli
Suitable host strains: DH5α, Fusion Blue, and other general purpose strains.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
E. coli replication origin: ColE1
Copy number: low
载体序列
LOCUS pRetroQ-DsRed-Monomer-N1 7571 bp DNA SYN
DEFINITION pRetroQ-DsRed-Monomer-N1
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7571
/organism="pRetroQ-DsRed-Monomer-N1"
/mol_type="other DNA"
promoter 2..506
/label="CMV_immearly_promoter"
misc_feature 6..688
/label="5_LTR"
misc_feature 9..296
/label="CAG_enhancer"
misc_feature 463..483
/label="CMV_fwd_primer"
misc_feature 758..1567
/label="psi_plus_pack"
misc_feature 1154..1561
/label="gag"
misc_feature 1497..1519
/label="MSCV_primer"
misc_feature 1497..1519
/label="pLXSN_5_primer"
misc_feature 1535..1551
/label="pBABE_5_primer"
promoter 1591..2143
/label="CMV_immearly_promoter"
misc_feature 1646..1933
/label="CAG_enhancer"
misc_feature 2100..2120
/label="CMV_fwd_primer"
promoter 2101..2170
/label="CMV_promoter"
CDS 2260..2937
/label="ORF frame 1"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA
KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG
GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE
ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE
ARHSGSQ*"
gene 2263..2934
/label="DsRed_Monomer"
/gene="DsRed_Monomer"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA
KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG
GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE
ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE
ARHSGSQ*"
misc_feature complement(2440..2460)
/label="dsRed1_N_primer"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA
KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG
GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE
ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE
ARHSGSQ*"
misc_feature complement(3001..3024)
/label="MSCV_rev_primer"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTA
KLQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDG
GVVEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGE
ISHALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAE
ARHSGSQ*"
CDS 3246..4085
/label="ORF frame 3"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
PGA*"
misc_feature 3398..3417
/label="mPGK_F_primer"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
PGA*"
CDS complement(3411..4136)
/label="ORF frame 1"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
gene 3486..4085
/label="puro (variant)"
/gene="puro (variant)"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(4805..4827)
/label="pGEX_3_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(4963..4983)
/label="pBABE_3_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature complement(4969..5184)
/label="SV40_enhancer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
promoter 4981..5249
/label="SV40_promoter"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
rep_origin 5148..5225
/label="SV40_origin"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
misc_feature 5210..5229
/label="SV40pro_F_primer"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
gene complement(6349..7209)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLGLQVERPGDEEEENSAADVRF*"
CDS complement(6349..7209)
/label="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(7251..7279)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
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pRetroQ-DsRed-Monomer-N1载体
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