pLXSN 载体描述pLXSN contains elements derived from Moloney murine leukemia virus (MoMuLV) and Moloney murine sarcoma virus (MoMuSV), and is designed for retroviral gene delivery and expression (1–3). Upon transfection into a packaging cell line, pLXSN can transiently express, or integrate and stably express, a transcript containing Ψ+ (the extended viral packaging signal) the gene of interest, and a selectable marker. The 5’ viral LTR in this vector contains promoter/ enhancer sequences that control expression of the gene of interest in the multiple cloning site. The SV40 early promoter (PSV40e) controls expression of the neomycin resistance gene (Neor), which allows antibiotic selection in eukaryotic cells. pLXSN also includes the Col E1 origin of replication and E. coli Ampr gene for propagation and antibiotic selection in bacteria.As a part of the Retro-X System (Cat. No. 631508), pLXSN can be transfected into a packaging cell line such as the RetroPack PT67 Cell Line (Cat. No. 631510). Once in the cell, RNA from the vector is packaged into infectious, replication-incompetent retroviral particles. pLXSN does not contain the structural genes (gag, pol, and env) necessary for particle formation and replication, however, these genes are stably integrated into PT67 (4–7).Subsequent introduction of pLXSN, containing Ψ+ (psi), transcription and processing elements, and the gene of interest produces high-titer, replication-incompetent infectious virus. That is, these retroviral particles can infect target cells and transmit the gene of interest (which is cloned between the viral LTR sequences), but cannot replicate within these cells since the cells lack the viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chances of producing replication-competent virus due to recombination events during cell proliferation.
LOCUS pLXSN 5874 bp DNA SYNDEFINITION pLXSNACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.FEATURES Location/Qualifiers source 1..5874 /organism="pLXSN" /mol_type="other DNA" misc_feature 175..763 /label="5MoMuLV_LTR" misc_feature 833..1642 /label="psi_plus_pack" misc_feature 1229..1636 /label="gag" misc_feature 1572..1594 /label="MSCV_primer" misc_feature 1572..1594 /label="pLXSN_5_primer" misc_feature 1610..1626 /label="pBABE_5_primer" misc_feature complement(1672..1692) /label="pBABE_3_primer" misc_feature complement(1678..1893) /label="SV40_enhancer" promoter 1690..1958 /label="SV40_promoter" rep_origin 1857..1934 /label="SV40_origin" misc_feature 1919..1938 /label="SV40pro_F_primer" CDS 2066..2860 /label="ORF frame 2" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" gene 2069..2857 /label="NeoR/KanR" /gene="NeoR/KanR" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" misc_feature 2920..3513 /label="3MoMuLV_LTR" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" misc_feature 2962..3513 /label="5_LTR2" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" misc_feature complement(3616..3638) /label="pGEX_3_primer" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" gene complement(4809..5669) /label="Ampicillin" /gene="Ampicillin" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" CDS complement(4809..5669) /label="ORF frame 2" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*" promoter complement(5711..5739) /label="AmpR_promoter" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*"ORIGIN 1 gaattgctag caattgctag caattgctag caattcatac cagatcaccg aaaactgtcc 61 tccaaatgtg tccccctcac actcccaaat tcgcgggctt ctgcctctta gaccactcta 121 ccctattccc cacactcacc ggagccaaag ccgcggccct tccgtttctt tgcttttgaa 181 agaccccacc cgtaggtggc aagctagctt aagtaacgcc actttgcaag gcatggaaaa 241 atacataact gagaatagaa aagttcagat caaggtcagg aacaaagaaa cagctgaata 301 ccaaacagga tatctgtggt aagcggttcc tgccccggct cagggccaag aacagatgag 361 acagctgagt gatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcggg 421 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag tgaatcatca 481 gatgtttcca gggtgcccca aggacctgaa aatgaccctg taccttattt gaactaacca 541 atcagttcgc ttctcgcttc tgttcgcgcg cttccgctct ccgagctcaa taaaagagcc 601 cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat 661 tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg 721 tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc tcgtccggga 781 tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac 841 ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta 901 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac 961 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac 1021 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga 1081 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag 1141 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt 1201 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt 1261 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag 1321 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga 1381 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc 1441 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc 1501 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc 1561 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc 1621 cctcactcct tctctaggcg ccggaattcg ttaactcgag gatccggctg tggaatgtgt 1681 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 1741 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 1801 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 1861 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 1921 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct 1981 tttttggagg cctaggcttt tgcaaaaagc ttgggctgca ggtcgaggcg gatctgatca 2041 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 2101 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 2161 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 2221 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 2281 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 2341 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 2401 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 2461 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 2521 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 2581 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 2641 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 2701 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 2761 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 2821 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgat 2881 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag 2941 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag 3001 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag 3061 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga 3121 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac 3181 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc 3241 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc 3301 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct 3361 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa 3421 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag 3481 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc 3541 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg 3601 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 3661 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 3721 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg 3781 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc 3841 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc 3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 3961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 4261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 4381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 4441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 4501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 4561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 4621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 4681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 4801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 4861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 4921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 4981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5101 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg 5161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 5341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 5401 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta 5461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 5521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 5581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 5641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 5701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5761 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 5821 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaa