质粒名称: | pLXRN |
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出品公司: | Addgene |
目录编号: | 99206 |
存储实验室: | Peter Pedersen |
载体骨架基本信息 |
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载体名称: | pLXSN |
载体出品公司: | Dr. Dusty Miller, Fred Hutchinson Cancer Center, Seattle, WA |
载体抗性: | 氨苄青霉素 |
载体修饰: | 将SV40启动子驱动的新霉素选择标记物替换为脑心肌炎病毒的内部核糖体进入位点/序列(IRES),生成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(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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启动子 | LTR |
克隆方法: | 限制性内切酶 |
5' 测序引物 | CCCTTGAACCTCCTCGTTCGACC |
3' 测序引物 | GTAAAGCATGTGCACCGAGGCC |
文献引用方法 |
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材料和方法部分: | pLXRN was a gift from Peter Pedersen (Addgene plasmid # 99206 ; http://n2t.net/addgene:99206 ; RRID:Addgene_99206) |
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 6091 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Bicistronic retroviral vector. ACCESSION . VERSION . KEYWORDS pLXRN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6091) TITLE pLXRN bicistronic retroviral vector JOURNAL Unpublished REFERENCE 2 (bases 1 to 6091) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6091 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 11..29 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" LTR 631..806 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 869..1068 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1269..1685 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1615..1637 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1653..1669 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1710..2281 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1875..1892) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2102..2121 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2317..3120 /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(2380..2399) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2990..3009 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 3180..3773 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(3876..3898) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3984..4124 /label=bom /note="basis of mobility region from pBR322" primer_bind 4039..4058 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4139..4156) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4310..4898) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4390..4409) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5069..5929) /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 5692..5711 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5930..6034) /gene="bla" /label=AmpR promoter ORIGIN 1 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattgc tagcaattgc 61 tagcaattgc tagcaattca taccagatca ccgaaaactg tcctccaaat gtgtccccct 121 cacactccca aattcgcggg cttctgcctc ttagaccact ctaccctatt ccccacactc 181 accggagcca aagccgcggc ccttccgttt ctttgctttt gaaagacccc acccgtaggt 241 ggcaagctag cttaagtaac gccactttgc aaggcatgga aaaatacata actgagaata 301 gaaaagttca gatcaaggtc aggaacaaag aaacagctga ataccaaaca ggatatctgt 361 ggtaagcggt tcctgccccg gctcagggcc aagaacagat gagacagctg agtgatgggc 421 caaacaggat atctgtggta agcagttcct gccccggctc ggggccaaga acagatggtc 481 cccagatgcg gtccagccct cagcagtttc tagtgaatca tcagatgttt ccagggtgcc 541 ccaaggacct gaaaatgacc ctgtacctta tttgaactaa ccaatcagtt cgcttctcgc 601 ttctgttcgc gcgcttccgc tctccgagct caataaaaga gcccacaacc cctcactcgg 661 cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt 721 gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg 781 actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag acccctgccc 841 agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg 901 attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta gctaactagc 961 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg 1021 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga 1081 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt 1141 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc 1201 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg 1261 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg 1321 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct 1381 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag 1441 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc 1501 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc 1561 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg 1621 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag 1681 gcgccggaat tcgttaactc gaggatccgc ccctctccct ccccccccta acgttactgg 1741 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1801 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1861 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1921 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1981 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 2041 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 2101 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 2161 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 2221 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata 2281 agcttgccac aacccgggat aattcctgca gccaatatgg gatcggccat tgaacaagat 2341 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 2401 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 2461 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 2521 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 2581 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 2641 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 2701 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 2761 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 2821 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga tgatctcgtc 2881 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 2941 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3001 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3061 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3121 gcgggactct ggggttcgat aaaataaaag attttattta gtctccagaa aaagggggga 3181 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca 3241 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca 3301 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca 3361 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc 3421 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta 3481 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt 3541 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa 3601 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg 3661 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc 3721 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc 3781 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct 3841 ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 3901 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 3961 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg 4021 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt 4081 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct 4141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4741 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 4861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 4921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 4981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5101 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5161 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5221 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5281 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5341 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt 5401 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5461 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 5521 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5581 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5641 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg 5701 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 5761 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5821 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 5881 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 5941 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6001 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6061 acgtctaaga aaccattatt atcatgacat t //
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