| 质粒名称: | pLenti CMV GFP Zeo (637-7) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 17449 |
| 存储实验室: | Eric Campeau, Paul Kaufman |
|
载体骨架基本信息 |
|
| 载体名称: | p156RRL-sinPPT-CMV-GFP-PRE/Nhe I |
| 载体出品公司: | -- |
| 载体抗性: | 博莱霉素 |
| 空载体大小: | 5728 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 筛选标记: | 博莱霉素 |
| 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: | GFP,enhanced GFP |
| 物种来源: | -- |
| 插入大小: | 719 bp |
| 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: | BamHI(未破坏) |
| 3' 克隆位点: | Sal I(未破坏) |
| 5' 测序引物 | CMVforw |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti CMV GFP Zeo (637-7) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 17449 ; http://n2t.net/addgene:17449 ; RRID:Addgene_17449) |
| 参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pLenti CMV GFP Zeo (637-7) 6474 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 3rd gen lentiviral eGFP expression vector, CMV promoter, Zeo.
ACCESSION .
VERSION .
KEYWORDS pLenti CMV GFP Zeo (637-7)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6474)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
REFERENCE 2 (bases 1 to 6474)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6474
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 71..659
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 560..579
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 813..830
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 947..968
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 983..1013
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1021..1037
/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 1026..1048
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 1045..1061
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1045..1061
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 1080..1100
/label=T3
/note="T3 promoter, forward primer"
promoter 1082..1100
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1128..1354
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 1355..1535
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1582..1707
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2200..2433
/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 2929..3045
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 3068..3371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3372..3575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3526..3546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 3572..3596
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 3665..3674
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3671..4390
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3716..3737)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3977..3996)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4324..4345
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 4406..4994
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4459..4479)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4877..4888)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5045..5065)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5050..5379
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5230..5365
/label=SV40 ori
/note="SV40 origin of replication"
promoter 5391..5438
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 5457..5831
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
LTR 5924..6157
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6229..6350
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6266..6285)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6320..6339
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
61 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
781 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
841 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
901 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
961 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
1021 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
1081 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
1141 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
1201 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
1261 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
1321 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
1381 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
1441 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
1501 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
1561 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
1621 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
1681 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
1741 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
1801 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
1861 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
1921 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
1981 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
2041 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
2101 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
2161 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
2221 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
2281 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
2341 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
2401 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
2461 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
2521 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
2581 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
2641 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
2701 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
2761 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
2821 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
2881 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
2941 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
3001 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg
3061 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
3121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
3181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
3241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
3301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
3361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
3421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
3481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
3541 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
3601 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctag aggatccacc
3661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
3721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
3781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
3841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
3901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
3961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
4021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
4081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
4141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
4201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
4261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
4321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
4381 gtacaagtaa agcggccgcg tcgacaatca acctctggat tacaaaattt gtgaaagatt
4441 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
4501 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
4561 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
4621 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
4681 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
4741 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
4801 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
4861 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
4921 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
4981 ggccgcctcc ccgcctggaa ttctgcagat atccggttag taatgagttt ggaattaatt
5041 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
5101 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
5161 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
5281 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
5341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccct gttgacaatt
5401 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
5461 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
5521 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
5581 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
5641 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
5701 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
5761 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
5821 agcaggactg acacgtgcta cgagatttaa atggtacctt taagaccaat gacttacaag
5881 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac
5941 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
6001 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
6061 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
6121 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
6181 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
6241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
6301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
6361 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
6421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccg
//
