| 质粒名称: | pLenti.PGK.chFP.W |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51008 |
| 存储实验室: | Rusty Lansford |
|
载体骨架基本信息 |
|
| 载体名称: | pRRLsin.cPPT.PGKp.eGFP.W |
| 载体出品公司: | L Naldini |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 7375 bp |
| 载体修饰: | pLenti.pgk：chFP慢病毒载体衍生自pRRLsin.cPPT.PGKp.eGFP.W慢病毒载体，其包含组成型表达的人磷酸甘油酸激酶启动子（De Palma等，2003）。 |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mChFP,mCherry FP |
| 物种来源: | -- |
| 插入大小: | 711 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | human PGK |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH1(未破坏) |
| 3' 克隆位点: | Sal11(未破坏) |
| 5' 测序引物 | T3 |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti.PGK.chFP.W was a gift from Rusty Lansford (Addgene plasmid # 51008 ; http://n2t.net/addgene:51008 ; RRID:Addgene_51008) |
买家导航
pLenti.PGK.chFP.W载体质粒基本信息
pLenti.PGK.chFP.W质粒图谱载体图谱和pLenti.PGK.chFP.W载体序列质粒序列多克隆位点信息
pLenti.PGK.chFP.W质粒载体简介
pLenti.PGK.chFP.W质粒序列载体序列
LOCUS pLenti.PGK.chFP.W 7363 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Lentivector that expresses chFP from internal human PGK promoter.
ACCESSION .
VERSION .
KEYWORDS pLenti.PGK.chFP.W
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7363)
TITLE Lentivirus
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7363)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7363
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 236..416
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 460..585
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1078..1311
/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 1807..1924
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1971..2481
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2400..2418
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 2491..3201
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2636..2654)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2904..2923
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 3217..3805
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3270..3290)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3688..3699)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3892..4125
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4197..4318
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4234..4253)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4288..4307
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4337..4472
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4399..4418
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4504..4523)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4505..4523)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4533..4550)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4533..4549)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4542..4564)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4694..5149
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4781..4800)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4991..5012
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5178..5282
/gene="bla"
/label=AmpR promoter
CDS 5283..6143
/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(5501..5520)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6314..6902
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6803..6822
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7056..7073
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7190..7211
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7226..7256
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7264..7280
/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 7269..7291
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7288..7304
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7288..7304
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7323..7343
/label=T3
/note="T3 promoter, forward primer"
promoter 7325..7343
/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 ctcgatacaa taaacgggtc
241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg
481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattcccac ggggttgggg
1981 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
2041 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
2101 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
2161 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
2221 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
2281 cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag cggccgggaa
2341 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
2401 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
2461 atcaccgacc tctctcccca gggggatccg atggtgagca agggcgagga ggataacatg
2521 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
2581 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
2641 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
2701 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
2761 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
2821 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
2881 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
2941 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
3001 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
3061 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
3121 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
3181 atggacgagc tgtacaagta aagcggccgc gtcgacaatc aacctctgga ttacaaaatt
3241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
3301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
3361 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
3421 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
3481 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
3541 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
3601 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
3661 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
3721 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
3781 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga
3841 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
3901 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
3961 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
4021 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
4081 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
4141 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
4201 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
4261 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
4321 atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcgtcgaga cgtacccaat
4501 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
4561 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
4621 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
4681 ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
4741 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
4801 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
4861 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
4921 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
4981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
5041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
5101 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ccaggtggca
5161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
5221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
5281 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
5341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
5401 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
5461 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
5521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
5581 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
5641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
5701 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
5761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
5821 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
5881 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
5941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
6001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
6061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
6121 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
6181 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
6241 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
6301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
6361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
6421 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
6481 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
6541 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
6601 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
6661 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
6721 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
6781 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
6841 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
6901 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
6961 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
7021 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
7081 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
7141 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
7201 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
7261 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
7321 gcgcaattaa ccctcactaa agggaacaaa agctggagct gca
//
