pLXRN

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载体基本信息

质粒名称: pLXRN 
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
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
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
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)

载体图谱质粒图谱和多克隆位点信息

pLXRN质粒图谱

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  JOURNAL   Unpublished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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