基本信息
| 启动子: | PGK ,CMV |
| 复制子: | pUC ori |
| 终止子: | SV40 poly(A) signal |
| 质粒分类: | 病毒系列，慢病毒克隆载体 |
| 质粒大小: | 8751bp |
| 原核抗性: | Amp |
| 筛选标记: | Puro |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37℃，有氧 LB |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 无须诱导，瞬时表达 |
| 5'测序引物: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
| 3'测序引物: | 根据序列设计引物 |
质粒简介
plvx-dsred-monomer-n1是HIV-1，慢病毒表达载体，允许你表达你的感兴趣的基因融合，DsRed Monomer，一个单体的突变红色荧光蛋白Discosoma sp.。基因克隆到多克隆位点（MCS），位于单体的基因编码序列上游，表示为N的单体蛋白融合蛋白。融合蛋白的表达是由组成性激活人类巨细胞病毒立即早期启动子（CMV）位于只是上游的MCS。慢病毒颗粒来自矢量允许表达红色荧光蛋白的单体融合蛋白在几乎任何类型的细胞，包括原代细胞。这个DsRed Monomer表示，未改性的载体，可用于生产标记病毒优化感染协议。
plvx-dsred-monomer-n1包含所有必要的病毒处理单元复制无能的病毒生产，以及元素来提高病毒滴度，转基因表达和整体载体功能。土拨鼠肝炎病毒转录后调节元件（WPRE）促进RNA处理事件提高病毒和转基因RNA的核出口，导致病毒滴度增加包装细胞，并增强你的基因在靶细胞中的表达。此外,载体包括Rev应答元件（RRE），从而进一步增加病毒滴度提高拼接的病毒RNA转运出细胞核。，pLVX DsRed—monomer-n1还包含一个ZY聚嘌呤道（CPPT）元，增加核在靶细胞感染过程中输入病毒基因组，导致改进的载体整合与更有效的转导。
质粒图谱
质粒序列
LOCUS Exported 8751 bp ds-DNA circular SYN 07-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8751)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, September 7, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..8751
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2793..2873
/note="MCS"
/note="multiple cloning site"
CDS 2881..3558
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Strongin et al., 2007)"
/note="DsRed-Monomer"
/note="mammalian codon-optimized"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
GSQ"
promoter 3587..4086
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4107..4706
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4720..5308
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5191..5202)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5515..6148
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6276..6292)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6300..6316
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6324..6354)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 6369..6390
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6678..7266)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7437..8297)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8298..8402)
/gene="bla"
/note="AmpR promoter"
polyA_signal 8450..8584
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc
2881 atggacaaca ccgaggacgt catcaaggag ttcatgcagt tcaaggtgcg catggagggc
2941 tccgtgaacg gccactactt cgagatcgag ggcgagggcg agggcaagcc ctacgagggc
3001 acccagaccg ccaagctgca ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
3061 ctgtcccccc agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc
3121 gactacatga agctgtcctt ccccgagggc ttcacctggg agcgctccat gaacttcgag
3181 gacggcggcg tggtggaggt gcagcaggac tcctccctgc aggacggcac cttcatctac
3241 aaggtgaagt tcaagggcgt gaacttcccc gccgacggcc ccgtaatgca gaagaagact
3301 gccggctggg agccctccac cgagaagctg tacccccagg acggcgtgct gaagggcgag
3361 atctcccacg ccctgaagct gaaggacggc ggccactaca cctgcgactt caagaccgtg
3421 tacaaggcca agaagcccgt gcagctgccc ggcaaccact acgtggactc caagctggac
3481 atcaccaacc acaacgagga ctacaccgtg gtggagcagt acgagcacgc cgaggcccgc
3541 cactccggct cccagtagag cggccgcgac tctagataat tctaccgggt aggggaggcg
3601 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
3661 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
3721 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
3781 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
3841 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
3901 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
3961 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
4021 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
4081 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
4141 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
4201 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
4261 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
4321 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
4381 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
4441 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
4501 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
4561 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
4621 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
4681 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa
4741 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
4801 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
4861 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
4921 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
4981 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
5041 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
5101 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
5161 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
5221 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
5281 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
5341 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
5401 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
5461 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
5521 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
5581 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
5641 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
5701 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
5761 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
5821 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
5881 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
5941 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
6001 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
6061 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
6121 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
6181 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta
6241 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
6421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
6481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
6541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
6601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
6841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
7081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
7201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
7261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
7321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
7381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
7441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
7501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
7561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
7621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
7681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
7741 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
7801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
7861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
7921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
7981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
8041 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
8101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
8161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
8221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
8281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
8341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
8401 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
8461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
8521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
8581 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
8641 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
8701 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
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