| 质粒名称: | pRRLSIN.cPPT.PGK-GFP.WPRE |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12252 |
| 存储实验室: | Didier Trono |
|
载体骨架基本信息 |
|
| 载体名称: | pRRL |
| 载体出品公司: | Naldini lab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,Lentiviral |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | See map |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRRLSIN.cPPT.PGK-GFP.WPRE was a gift from Didier Trono (Addgene plasmid # 12252 ; http://n2t.net/addgene:12252 ; RRID:Addgene_12252) |
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 pRRLSIN.cPPT.PGK-GFP.WPRE 7388 bp ds-DNA circular SYN 13-SEP-2018
DEFINITION 3rd generation lentiviral backbone.
ACCESSION .
VERSION .
KEYWORDS pRRLSIN.cPPT.PGK-GFP.WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7388)
TITLE Trono Lab Misc Plasmids
REFERENCE 2 (bases 1 to 7388)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 13, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7388
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 8..234
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 235..415
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 462..587
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1080..1313
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1809..1926
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1972..2482
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2401..2419
/label=hPGK-F
/note="Human PGK promoter, forward primer"
regulatory 2498..2507
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2504..3223
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2549..2570)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2810..2829)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3157..3178
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3239..3827
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3292..3312)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3710..3721)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3914..4147
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4219..4340
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4256..4275)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4310..4329
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4380..4515
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4442..4461
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4535..4554)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4536..4554)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4564..4581)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4564..4580)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4573..4595)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4722..5177
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4809..4828)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5019..5040
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5203..5307
/gene="bla"
/label=AmpR promoter
CDS 5308..6168
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5526..5545)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6339..6927
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6828..6847
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7081..7098
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7215..7236
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7251..7281
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7289..7305
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
primer_bind 7294..7316
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7313..7329
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7313..7329
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7348..7368
/label=T3
/note="T3 promoter, forward primer"
promoter 7350..7368
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
1921 aattttatcg atcacgagac tagcctcgag aagcttgata tcgaattcca cggggttggg
1981 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
2041 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
2101 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
2161 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
2221 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
2281 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
2341 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
2401 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
2461 aatcaccgac ctctctcccc agggggatcc accggtcgcc accatggtga gcaagggcga
2521 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
2581 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
2641 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
2701 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
2761 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
2821 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
2881 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
2941 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
3001 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
3061 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
3121 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
3181 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgtcgacaa
3241 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
3301 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
3361 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
3421 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
3481 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
3541 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
3601 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
3661 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
3721 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
3781 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
3841 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
3901 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
3961 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
4021 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
4081 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
4141 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
4201 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
4261 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
4321 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
4441 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
4501 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
4561 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
4621 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
4681 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
4741 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
4801 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
4861 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
4921 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
4981 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
5041 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
5101 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
5161 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
5221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
5281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
5341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
5401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
5461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
5521 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
5581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
5641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
5701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
5761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
5821 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
5881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
5941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
6001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
6061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
6121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
6181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
6241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
6301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
6361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
6421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
6481 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
6541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
6601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
6661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
6721 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
6781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
6841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
6901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
6961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
7021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
7081 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
7141 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
7201 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
7261 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
7321 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
7381 gagctgca
//
