| 质粒名称: | pVITRO1-SS-113 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 68737 |
| 存储实验室: | James Collins |
|
载体骨架基本信息 |
|
| 载体名称: | pVITRO1-NEO-MCS |
| 载体出品公司: | invivogen |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 6295 bp |
| 完整质粒大小: | 7477 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | ZF9-VP64 |
| 物种来源: | Synthetic |
| 插入大小: | 486 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
flag（插入N端子）
NLS（插入件上的N端子）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pVITRO1-SS-113 was a gift from James Collins (Addgene plasmid # 68737 ; http://n2t.net/addgene:68737 ; RRID:Addgene_68737) |
| 参考文献部分: | DNA sense-and-respond protein modules for mammalian cells. Slomovic S, Collins JJ. Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585. 10.1038/nmeth.3585 PubMed 26389572 |
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 pVITRO1-SS-113 7477 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION ZF9-TF with mCherry marker.
ACCESSION .
VERSION .
KEYWORDS pVITRO1-SS-113
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7477)
AUTHORS Slomovic S, Collins JJ
TITLE DNA sense-and-respond protein modules for mammalian cells.
JOURNAL Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
PUBMED 26389572
REFERENCE 2 (bases 1 to 7477)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7477
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(74..94)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
intron 583..1530
/label=mEF-1-alpha intron
/note="intron upstream of the start codon of mouse
EF-1-alpha"
CDS 1585..1608
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1609..1629
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1909..2058
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
polyA_signal 2090..2211
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2101..2120)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2155..2174
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 2399..2987
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2888..2907
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 3067..3370
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
CDS 4803..5513
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWQASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(4948..4966)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 5216..5235
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 5520..5964
/label=FMDV IRES
/note="internal ribosome entry site (IRES) of the
foot-and-mouth disease virus"
promoter 5997..6043
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter 6035..6053
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 6063..6857
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6117..6136)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6727..6746
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6904..7476
/label=EF-1-alpha poly(A) signal
/note="human EF-1-alpha polyadenylation signal"
ORIGIN
1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
1561 aaccggtgat atcaaagatc tatggactac aaggacgacg atgacaagcc caagaaaaag
1621 cgcaaggtct ctagacccgg ggagcgcccc ttccagtgtc gcatttgcat gcggaacttt
1681 tcggataaaa ctaaattgag agttcatacc cgtactcata ccggtgaaaa accgtttcag
1741 tgtcggatct gtatgcgaaa tttctccgtt agacataatt tgactagaca tctacgtacg
1801 cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag tcaatctact
1861 tctttgcaaa gacacctaaa aacccacctg agaggatccg ggcgcgccga cgcgctggac
1921 gatttcgatc tcgacatgct gggttctgat gccctcgatg actttgacct ggatatgttg
1981 ggaagcgacg cattggatga ctttgatctg gacatgctcg gctccgatgc tctggacgat
2041 ttcgatctcg atatgttaat taattgttaa gctagctggc cagacatgat aagatacatt
2101 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
2161 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
2221 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
2281 taaaacctct acaaatgtgg tatggaaatg ttaattaact agccatgacc aaaatccctt
2341 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
2401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
2461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
2521 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
2581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
2641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
2701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
2761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
2821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
2881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
2941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
3001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc ttaattaacc
3061 tgcaggcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
3121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
3181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
3241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
3301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
3361 tattaccatg atgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
3421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt gactagtgga
3481 gccgagagta attcatacaa aaggagggat cgccttcgca aggggagagc ccagggaccg
3541 tccctaaatt ctcacagacc caaatccctg tagccgcccc acgacagcgc gaggagcatg
3601 cgcccagggc tgagcgcggg tagatcagag cacacaagct cacagtcccc ggcggtgggg
3661 ggaggggcgc gctgagcggg ggccagggag ctggcgcggg gcaaactggg aaagtggtgt
3721 cgtgtgctgg ctccgccctc ttcccgaggg tgggggagaa cggtatataa gtgcggtagt
3781 cgccttggac gttctttttc gcaacgggtt tgccgtcaga acgcaggtga gtggcgggtg
3841 tggcttccgc gggccccgga gctggagccc tgctctgagc gggccgggct gatatgcgag
3901 tgtcgtccgc agggtttagc tgtgagcatt cccacttcga gtggcgggcg gtgcgggggt
3961 gagagtgcga ggcctagcgg caaccccgta gcctcgcctc gtgtccggct tgaggcctag
4021 cgtggtgtcc gccgccgcgt gccactccgg ccgcactatg cgttttttgt ccttgctgcc
4081 ctcgattgcc ttccagcagc atgggctaac aaagggaggg tgtggggctc actcttaagg
4141 agcccatgaa gcttacgttg gataggaatg gaagggcagg aggggcgact ggggcccgcc
4201 cgccttcgga gcacatgtcc gacgccacct ggatggggcg aggcctgtgg ctttccgaag
4261 caatcgggcg tgagtttagc ctacctgggc catgtggccc tagcactggg cacggtctgg
4321 cctggcggtg ccgcgttccc ttgcctccca acaagggtga ggccgtcccg cccggcacca
4381 gttgcttgcg cggaaagatg gccgctcccg gggccctgtt gcaaggagct caaaatggag
4441 gacgcggcag cccggtggag cgggcgggtg agtcacccac acaaaggaag agggccttgc
4501 ccctcgccgg ccgctgcttc ctgtgacccc gtggtctatc ggccgcatag tcacctcggg
4561 cttctcttga gcaccgctcg tcgcggcggg gggaggggat ctaatggcgt tggagtttgt
4621 tcacatttgg tgggtggaga ctagtcaggc cagcctggcg ctggaagtca ttcttggaat
4681 ttgccccttt gagtttggag cgaggctaat tctcaagcct cttagcggtt caaaggtatt
4741 ttctaaaccc gtttccaggt gttgtgaaag ccaccgctaa ttcaaagcaa tccggagtat
4801 acatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
4861 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
4921 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
4981 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
5041 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
5101 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
5161 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
5221 taatgcagaa gaagaccatg ggctggcagg cctcctccga gcggatgtac cccgaggacg
5281 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
5341 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
5401 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
5461 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taacctagga
5521 gcaggtttcc ccaatgacac aaaacgtgca acttgaaact ccgcctggtc tttccaggtc
5581 tagaggggta acactttgta ctgcgtttgg ctccacgctc gatccactgg cgagtgttag
5641 taacagcact gttgcttcgt agcggagcat gacggccgtg ggaactcctc cttggtaaca
5701 aggacccacg gggccaaaag ccacgcccac acgggcccgt catgtgtgca accccagcac
5761 ggcgacttta ctgcgaaacc cactttaaag tgacattgaa actggtaccc acacactggt
5821 gacaggctaa ggatgccctt caggtacccc gaggtaacac gcgacactcg ggatctgaga
5881 aggggactgg ggcttctata aaagcgctcg gtttaaaaag cttctatgcc tgaataggtg
5941 accggaggtc ggcacctttc ctttgcaatt actgacccta tgaatacact gactgtttga
6001 caattaatca tcggcatagt atatcggcat agtataatac gactcactat aggagggcca
6061 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
6121 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
6181 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
6241 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
6301 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
6361 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
6421 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
6481 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
6541 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
6601 acggcgagga tctcgtcgtg acacatggcg atgcctgctt gccgaatatc atggtggaaa
6661 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
6721 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
6781 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
6841 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgaattcgct
6901 aggattatcc ctaatacctg ccaccccact cttaatcagt ggtggaagaa cggtctcaga
6961 actgtttgtt tcaattggcc atttaagttt agtagtaaaa gactggttaa tgataacaat
7021 gcatcgtaaa accttcagaa ggaaaggaga atgttttgtg gaccactttg gttttctttt
7081 ttgcgtgtgg cagttttaag ttattagttt ttaaaatcag tactttttaa tggaaacaac
7141 ttgaccaaaa atttgtcaca gaattttgag acccattaaa aaagttaaat gagaaacctg
7201 tgtgttcctt tggtcaacac cgagacattt aggtgaaaga catctaattc tggttttacg
7261 aatctggaaa cttcttgaaa atgtaattct tgagttaaca cttctgggtg gagaataggg
7321 ttgttttccc cccacataat tggaagggga aggaatatca tttaaagcta tgggagggtt
7381 gctttgatta caacactgga gagaaatgca gcatgttgct gattgcctgt cactaaaaca
7441 ggccaaaaac tgagtccttg ggttgcatag aaagctg
//
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