| 质粒名称: | pLXRN-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99207 |
| 存储实验室: | Peter Pedersen |
|
载体骨架基本信息 |
|
| 载体名称: | pLXSN |
| 载体出品公司: | 华盛顿州西雅图弗雷德·哈钦森癌症中心的达斯提·米勒博士 |
| 载体抗性: | 氨苄西林 |
| 载体修饰: | SV40启动子驱动的新霉素选择标记被脑心肌炎病毒的内部核糖体入口位点/序列（IRES）替换，以产生双顺反子逆转录病毒载体。 |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 新霉素（用G418选择） |
| 高拷贝/低拷贝: | 高拷贝 |
| 宿生菌： | NEB Stable |
| 生长说明： | 可以在大肠杆菌dh5alpha中繁殖。大肠杆菌确定或大肠杆菌stbl建议减少重组事件 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 备用名称： | 增强型绿色荧光蛋白 |
| 物种来源: | 合成纤维；维多利亚绿松花 |
| 插入大小: | 715bp |
| 基因突变: | 结合Kozak序列来提高翻译效率。 |
| GenBank编号： | U55762 |
| 启动子： | CMV |
| 克隆方法: | 限制性酶 |
| 5' 克隆位点: | Ecori（未销毁） |
| 3' 克隆位点: | XHOI（未销毁） |
| 5' 测序引物 |
CCCTTGAACCTCCTCGTTCGACC
|
| 3' 测序引物 | CCTCACATTGCCAAAAGACG |
|
文献引用方法 |
|
| 材料和方法部分: | pLXRN-EGFP was a gift from Peter Pedersen (Addgene plasmid # 99207 ; http://n2t.net/addgene:99207 ; RRID:Addgene_99207) |
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
载体简介
载体序列
LOCUS pLXRN-EGFP 6812 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION EGFP expressing bicistronic retroviral vector.
ACCESSION .
VERSION .
KEYWORDS pLXRN-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6812)
TITLE pLXRN bicistronic retroviral vector
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6812)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6812
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 163..184
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 242..813
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(407..424)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 634..653
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 849..1652
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(912..931)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1522..1541
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
LTR 1712..2305
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(2408..2430)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2516..2656
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2571..2590
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2671..2688)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2842..3430)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2922..2941)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3601..4461)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4224..4243
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4462..4566)
/gene="bla"
/label=AmpR promoter
primer_bind 4634..4652
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR 5254..5429
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 5492..5691
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 5892..6308
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 6238..6260
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 6276..6292
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 6316..6325
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6322..229
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6367..6388)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6628..6647)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
ORIGIN
1 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
61 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
121 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
181 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac tcgaggatcc
241 gcccctctcc ctcccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
301 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
361 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
421 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
481 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
541 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
601 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
661 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
721 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
781 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccggg ataattcctg
841 cagccaatat gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt
901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
1021 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
1081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
1141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
1201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
1261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
1321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
1381 aggcgcgcat gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga
1441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
1501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
1561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
1621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg ataaaataaa
1681 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
1741 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
1801 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
1861 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
1921 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
1981 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
2041 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
2101 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
2161 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
2221 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
2281 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
2341 gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg
2401 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
2461 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
2521 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
2581 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
2641 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
3241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
3661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
3901 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
4201 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
4561 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
4621 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tgctagcaat
4681 tgctagcaat tgctagcaat tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc
4741 cctcacactc ccaaattcgc gggcttctgc ctcttagacc actctaccct attccccaca
4801 ctcaccggag ccaaagccgc ggcccttccg tttctttgct tttgaaagac cccacccgta
4861 ggtggcaagc tagcttaagt aacgccactt tgcaaggcat ggaaaaatac ataactgaga
4921 atagaaaagt tcagatcaag gtcaggaaca aagaaacagc tgaataccaa acaggatatc
4981 tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca gatgagacag ctgagtgatg
5041 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcggggcca agaacagatg
5101 gtccccagat gcggtccagc cctcagcagt ttctagtgaa tcatcagatg tttccagggt
5161 gccccaagga cctgaaaatg accctgtacc ttatttgaac taaccaatca gttcgcttct
5221 cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa agagcccaca acccctcact
5281 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
5341 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
5401 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
5461 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
5521 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
5581 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
5641 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
5701 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
5761 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
5821 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
5881 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
5941 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
6001 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
6061 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
6121 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
6181 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
6241 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
6301 taggcgccgg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
6361 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
6421 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
6481 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
6541 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
6601 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
6661 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
6721 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
6781 atggccgaca agcagaagaa cggcatcaag gt
//
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