| 质粒名称: | pMSCV-IRES-DsRed FP (pMIDsRed II) |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 52110 |
| 存储实验室: | Dario Vignali |
|
载体骨架基本信息 |
|
| 载体名称: | MSCV IRES GFP |
| 载体修饰: | expanded MCS and switched FP |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | IRES-DsRed (C terminal on backbone) |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | pLXSN_5_primer |
|
文献引用方法 |
|
| 材料和方法部分: | pMSCV-IRES-DsRed FP (pMIDsRed II) was a gift from Dario Vignali (Addgene plasmid # 52110 ; http://n2t.net/addgene:52110 ; RRID:Addgene_52110) |
| 参考文献部分: | Generation of T-cell receptor retrogenic mice. Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ, Vignali DA. Nat Protoc. 2006;1(1):406-17. 10.1038/nprot.2006.61 PubMed 17406263 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
52110-质粒图谱.png)
载体简介
载体序列
LOCUS pMSCV-IRES-DsRed FP (pMIDsRed II) 6468 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION a bicistronic IRES-DsRed containing retroviral vector, MCS is
expanded from pMSCV-IRES-GFP.
ACCESSION .
VERSION .
KEYWORDS pMSCV-IRES-DsRed FP (pMIDsRed II)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6468)
AUTHORS Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ,
Vignali DA
TITLE Generation of T-cell receptor retrogenic mice.
JOURNAL Nat Protoc. 2006;1(1):406-17.
PUBMED 17406263
REFERENCE 2 (bases 1 to 6468)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6468
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 169..510
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 577..993
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 923..945
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 961..977
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(1206..1223)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1433..1452
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1614..2291
/codon_start=1
/product="rapidly maturing tetrameric variant of DsRed
fluorescent protein (Bevis and Glick, 2002)"
/label=DsRed-Express
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
LFL"
primer_bind complement(1744..1762)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind complement(1794..1814)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 2012..2031
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2206..2229
/label=DsRed1-C
/note="DsRed1, forward primer"
LTR 2451..2965
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 3127..3143
/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(3151..3181)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3196..3217
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3334..3351)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3505..4093)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3585..3604)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4264..5124)
/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 4887..4906
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5125..5229)
/gene="bla"
/label=AmpR promoter
primer_bind 5297..5315
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5353..5375)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5475..5494
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5688..5710
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5838..5857
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 6057..105
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
ORIGIN
1 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
61 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
121 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
181 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
241 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
301 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
361 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
481 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
541 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
601 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
661 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
721 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
781 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
841 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
901 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
961 ctttatccag ccctcactcc ttctctaggc gccggaattc agatcttacg tagctagcgg
1021 atcccaattg ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg
1081 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
1141 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
1201 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
1261 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
1321 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
1381 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
1441 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
1501 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
1561 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggcct
1621 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
1681 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
1741 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
1801 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
1861 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
1921 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga
1981 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct
2041 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
2101 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg
2161 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
2221 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc
2281 tgttcctgta gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc
2341 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc
2401 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
2461 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
2521 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
2581 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
2641 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
2701 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
2761 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
2821 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
2881 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
2941 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
3001 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
3061 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
5221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
5341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
5401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
5461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
5521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
5581 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
5641 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
5701 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
5761 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
5821 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
5881 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac
5941 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata
6001 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
6061 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
6121 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga
6181 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
6241 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
6301 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
6361 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
6421 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgt
//
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