| 质粒名称: | pLXIN2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99203 |
| 存储实验室: | Saroj Mathupala |
|
载体骨架基本信息 |
|
| 载体名称: | pLXSN |
| 载体出品公司: | Miller，A.D.，Genbank，编号M28248 |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 5874 bp |
| 载体修饰: | 从EMCV（脑心肌炎病毒；Genbank NC甾0 479）中用IRES（内部核糖体进入位点/序列）替换驱动新霉素（遗传素；G-418）选择plxsn的内部SV40启动子，以产生这种双顺反式病毒载体。IRES保持不变，新霉素磷酸转移酶的翻译起始发生在IRES的第11个ATG，与野生型病毒相同。大多数商业质粒和逆转录病毒载体都有一个IRE，第11个ATG缺失。翻译起始于合成的ATG密码子下游19 bp处。PLXSN、Ecori Hpai Xhoi Bamhi的原始MCS保持不变。 |
| 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） |
| 生长条件: | 37 ℃ |
| 启动子: | LTR |
|
插入基因信息 |
|
| 克隆方法: | 限制性酶 |
| 5' 测序引物 | CCCTTGAACCTCCTCGTTCGACC |
| 3' 测序引物 | GTGCAATCCATCTTGTTCAATGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pLXIN2 was a gift from Saroj Mathupala (Addgene plasmid # 99203 ; http://n2t.net/addgene:99203 ; RRID:Addgene_99203) |
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 Exported 6057 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bicistronic retroviral vector (optimized internal ribosome entry
site).
ACCESSION .
VERSION .
KEYWORDS pLXIN2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6057)
TITLE pLXIN2
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6057)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6057
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 227..426
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 627..1043
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 973..995
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1011..1027
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 1068..1639
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 1089..1641
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1233..1250)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1460..1479
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1641..2444
/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(1704..1723)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2314..2333
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
LTR 2504..3097
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3200..3222)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3308..3448
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3363..3382
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3463..3480)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3634..4222)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3714..3733)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4393..5253)
/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 5016..5035
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5254..5358)
/gene="bla"
/label=AmpR promoter
primer_bind 5426..5444
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR 6046..164
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
ORIGIN
1 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
61 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
121 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
181 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
241 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
301 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
361 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
421 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
481 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
541 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
601 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
661 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
721 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
781 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
841 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
901 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
961 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
1021 ccctcactcc ttctctaggc gccggaattc gttaactcga ggatccgccc ctctccctcc
1081 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
1141 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
1201 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
1261 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaagcaa cgtctgtagc
1321 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
1381 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagtaggat
1441 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
1501 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
1561 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
1621 ttgaaaaaca cgatgataat atgggatcgg ccattgaaca agatggattg cacgcaggtt
1681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
1741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
1801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
1861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
1921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
1981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
2041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
2101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
2161 tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
2221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
2281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
2341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
2401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
2461 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
2521 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
2581 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
2641 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
2701 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
2761 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
2821 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
2881 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
2941 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
3001 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
3061 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
3121 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
3181 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
3241 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
3301 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
3361 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
3421 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
3481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
3541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
3601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
3661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
3721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
3781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
3841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
3901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
3961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
4021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
4081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
4141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
4201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
4261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
4321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
4381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
4441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
4501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
4561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
4621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
4681 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
4741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
4801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
4861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
4921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
4981 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
5041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
5101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
5161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
5221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
5281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
5341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
5401 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
5461 attgctagca attgctagca attgctagca attcatacca gatcaccgaa aactgtcctc
5521 caaatgtgtc cccctcacac tcccaaattc gcgggcttct gcctcttaga ccactctacc
5581 ctattcccca cactcaccgg agccaaagcc gcggcccttc cgtttctttg cttttgaaag
5641 accccacccg taggtggcaa gctagcttaa gtaacgccac tttgcaaggc atggaaaaat
5701 acataactga gaatagaaaa gttcagatca aggtcaggaa caaagaaaca gctgaatacc
5761 aaacaggata tctgtggtaa gcggttcctg ccccggctca gggccaagaa cagatgagac
5821 agctgagtga tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcggggc
5881 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagtg aatcatcaga
5941 tgtttccagg gtgccccaag gacctgaaaa tgaccctgta ccttatttga actaaccaat
6001 cagttcgctt ctcgcttctg ttcgcgcgct tccgctctcc gagctcaata aaagagc
//
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