| 质粒名称: | pLV-ER GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 80069 |
| 存储实验室: | Pantelis Tsoulfas |
|
载体骨架基本信息 |
|
| 载体名称: | pLV MP2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6782 bp |
| 完整质粒大小: | 7823 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 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: | ER GFP,AcGFP SEC61B,AcGFP Sec61 translocon beta subunit |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1041 bp |
| 基因突变: | -- |
| GenBank编号 | NM_006808 |
| 启动子 | CMV |
| 标签/融合蛋白: | 绿色荧光蛋白（GFP）来源于蓝绿豆，在NH2末端与SEC61B全长融合（插入物上的C末端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XhoI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLV-ER GFP was a gift from Pantelis Tsoulfas (Addgene plasmid # 80069 ; http://n2t.net/addgene:80069 ; RRID:Addgene_80069) |
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 pLV-ER GFP 7781 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION To express and target GFP into the endoplasmic reticulum using
lentivirus.
ACCESSION .
VERSION .
KEYWORDS pLV-ER GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7781)
TITLE pLV-ER eGFP
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7781)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7781
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 3..229
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 230..410
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 457..582
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1075..1308
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1493..1537
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 1804..1920
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 1943..2246
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2247..2450
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2401..2421
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2447..2471
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2551..3267
/codon_start=1
/product="Aequorea coerulescens GFP"
/label=AcGFP1
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
CDS 3283..3573
/codon_start=1
/gene="SEC61B"
/product="human Sec61-beta subunit of the translocon in the
endoplasmic reticulum"
/label=Sec61-beta
/translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
misc_feature 3627..4215
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3680..3700)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4098..4109)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4302..4535
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4607..4728
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4644..4663)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4698..4717
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4768..4903
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4830..4849
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4923..4942)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4924..4942)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4952..4969)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4952..4968)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4961..4983)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5110..5565
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5197..5216)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5407..5428
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5591..5695
/gene="bla"
/label=AmpR promoter
CDS 5696..6556
/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(5914..5933)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6727..7315
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7216..7235
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7469..7486
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7603..7624
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7639..7669
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7677..7693
/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 7682..7704
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7701..7717
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7701..7717
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7736..7756
/label=T3
/note="T3 promoter, forward primer"
promoter 7738..7756
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
1921 atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
1981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
2041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
2101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
2161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
2221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
2281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
2341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
2401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
2461 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccgcc
2521 cgggcgtacg ggatcccacc ggtgctcgag atggtgagca agggcgccga gctgttcacc
2581 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg
2641 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
2701 accggcaagc tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag
2761 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct
2821 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc
2881 gccgaggtga agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat
2941 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat
3001 gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac
3061 aacatcgagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc
3121 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag
3181 gaccccaacg agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc
3241 acccacggca tggatgagct gtacaagtcc ggactcagat ctatgcctgg tccgaccccc
3301 agtggcacta acgtgggatc ctcagggcgc tctcccagca aagcagtggc cgcccgggcg
3361 gcgggatcca ctgtccggca gaggaaaaat gccagctgtg ggacaaggag tgcaggccgc
3421 acaacctcgg caggcaccgg ggggatgtgg cgattctaca cagaagattc acctgggctc
3481 aaagttggcc ctgttccagt attggttatg agtcttctgt tcatcgcttc tgtatttatg
3541 ttgcacattt ggggcaagta cactcgttcg tagtctagac ctgcagggtc gactccggat
3601 gatcagggcc ctgtacagat atcgacaatc aacctctgga ttacaaaatt tgtgaaagat
3661 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
3721 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
3781 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
3841 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
3901 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
3961 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
4021 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
4081 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
4141 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
4201 gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga cttacaaggc
4261 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
4321 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
4381 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
4441 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
4501 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
4561 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
4621 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
4681 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
4741 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
4801 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
4861 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
4921 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
4981 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
5041 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
5101 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
5161 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
5221 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
5281 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
5341 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
5401 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
5461 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
5521 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
5581 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
5641 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
5701 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
5761 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
5821 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
5881 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
5941 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
6001 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
6061 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
6121 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
6181 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
6241 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
6301 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
6361 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
6421 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
6481 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
6541 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
6601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
6661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
6721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
6781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
6841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
6901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
6961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
7021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
7081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
7141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
7201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
7261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
7321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
7381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
7441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
7501 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
7561 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
7621 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
7681 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
7741 taaccctcac taaagggaac aaaagctgga gctgcaagct t
//
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