| 质粒名称: | pFlagCMV2-EFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12449 |
| 存储实验室: | Dong-Er Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pFlag-CMV2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4679bp |
| 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: | EFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pFlagCMV2-EFP was a gift from Dong-Er Zhang (Addgene plasmid # 12449 ; http://n2t.net/addgene:12449 ; RRID:Addgene_12449) |
| 参考文献部分: | The interferon-inducible ubiquitin-protein isopeptide ligase (E3) EFP also functions as an ISG15 E3 ligase. Zou W, Zhang DE. J Biol Chem. 2006 Feb 17. 281(7):3989-94. 10.1074/jbc.M510787200 PubMed 16352599 |
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 pFlagCMV2-EFP 7087 bp ds-DNA circular SYN 05-FEB-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFlagCMV2-EFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7087)
AUTHORS Zou W, Zhang DE
TITLE The interferon-inducible ubiquitin-protein isopeptide ligase (E3)
EFP also functions as an ISG15 E3 ligase.
JOURNAL J Biol Chem. 2006 Feb 17. 281(7):3989-94.
PUBMED 16352599
REFERENCE 2 (bases 1 to 7087)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 5, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7087
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 618..641
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 3111..3733
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3310..3329)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3757..3777)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3762..4091
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3942..4077
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4004..4023
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4129..4148)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4130..4148)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4162..4178)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4162..4178)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4175..4197)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4186..4202
/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)."
promoter complement(4210..4240)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4255..4276
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4393..4410)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4564..5152)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4644..4663)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5323..6183)
/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 5946..5965
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6184..6288)
/gene="bla"
/label=AmpR promoter
rep_origin 6315..6770
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6402..6421)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6612..6633
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6896..6918
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6910..6927
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6911..6927
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agaattgatc taccatggac tacaaagacg atgacgacaa gctcatggca gagctgtgcc
661 ccctggccga ggagctgtcg tgctccatct gcctggagcc cttcaaggag ccggtcacca
721 ctccgtgcgg ccacaacttc tgcgggtcgt gcctgaatga gacgtgggca gtccagggct
781 cgccatacct gtgcccgcag tgccgcgccg tctaccaggc gcgaccgcag ctgcacaaga
841 acacggtgct gtgcaacgtg gtggagcagt tcctgcaggc cgacctggcc cgggagccac
901 ccgccgacgt ctggacgccg cccgcccgcg cctctgcacc cagcccgaat gcccaggtgg
961 cctgcgacca ctgcctgaag gaggccgccg tgaagacgtg cttggtgtgc atggcctcct
1021 tctgtcagga gcacctgcag ccgcacttcg acagccccgc cttccaggac cacccgctgc
1081 agccgcccgt tcgcgacctg ttgcgccgca aatgttccca gcacaatcgg ctgcgggaat
1141 ttttctgccc cgagcacagc gagtgcatct gccacatctg cctggtggag cataagacct
1201 gctctcccgc gtccctgagc caggccagcg ccgacctgga ggccaccctg aggcacaaac
1261 taactgtcat gtacagtcag atcaacgggg cgtcgagagc actggatgat gtgagaaaca
1321 ggcagcagga tgtgcggatg actgcaaaca gaaaggtgga gcagctacaa caagaataca
1381 cggaaatgaa ggctctcttg gacgcctcag agaccacctc gacaaggaag ataaaggaag
1441 aggagaagag ggtcaacagc aagtttgaca ccatttatca gattctcctc aagaagaaga
1501 gtgagatcca gaccttgaag gaggagattg aacagagcct gaccaagagg gatgagttcg
1561 agtttctgga gaaagcatca aaactgcgag gaatctcaac aaagccagtc tacatccccg
1621 aggtggaact gaaccacaag ctgataaaag gcatccacca gagcaccata gacctcaaaa
1681 acgagctgaa gcagtgcatc gggcggctcc aggagctcac ccccagttca ggtgaccctg
1741 gagagcatga cccagcgtcc acacacaaat ccacacgccc tgtgaagaag gtctccaaag
1801 aggaaaagaa atccaagaaa cctccccctg tccctgcctt acccagcaag cttcccacgt
1861 ttggagcccc ggaacagtta gtggatttaa aacaagctgg cttggaggct gcagccaaag
1921 ccaccagctc acatccgaac tcaacatctc tcaaggccaa ggtgctggag accttcctgg
1981 ccaagtccag acctgagctc ctggagtatt acattaaagt catcctggac tacaacaccg
2041 cccacaacaa agtggctctg tcagagtgct atacagtagc ttctgtggct gagatgcctc
2101 agaactaccg gccgcatccc cagaggttca catactgctc tcaggtgctg ggcctgcact
2161 gctacaagaa ggggatccac tactgggagg tggagctgca gaagaacaac ttctgtgggg
2221 taggcatctg ctacggaagc atgaaccggc agggcccaga aagcaggctc ggccgcaaca
2281 gcgcctcctg gtgcgtggag tggttcaaca ccaagatctc tgcctggcac aataacgtgg
2341 agaaaaccct gccctccacc aaggccacgc gggtgggcgt gcttctcaac tgtgaccacg
2401 gctttgtcat cttcttcgct gttgccgaca aggtccacct gatgtataag ttcagggtgg
2461 actttactga ggctttgtac ccggctttct gggtattttc tgctggtgcc acactctcca
2521 tctgctcccc caagtaggca ggctgtaggc acttgggctg actgcctgca gaagtcccaa
2581 gaccctagtg aaaatacagc aggcagaact ctccttggat aattccccaa gaggtcccaa
2641 ggattgggag catgggaggg gagctggcgg gagggtggga ggtgggattt agccaggaaa
2701 ggggtgagag tgattgtgtt gtgggcgagg aggcgtttcc accccctggt gcctatcagg
2761 gcagggtgac ctactcccca ttgttctgga aatctccagg ctgctgggca gctgggcaga
2821 gctctgggaa gtgaagtcat gagtgcccga ttcctcttag agaaaatcca tagctactgt
2881 aggttctgtc ttgggccact tggatctgaa ggctgcccct ttgctctctg gggtagcctt
2941 cagatcttgg tgttttgaat tcttactata gatgttttta aagttccaaa gtcattgagt
3001 ttcaatgtta cataaactcc attacccgcn aaaaaaaaaa aaaaaaaaaa gggcggccgc
3061 gaattcatcg atagatctga tatcggtacc agtcgactct agaggatccc gggtggcatc
3121 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
3181 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
3241 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
3301 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
3361 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
3421 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
3481 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
3541 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
3601 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
3661 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
3721 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
3841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
3901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
3961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
4021 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
4081 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
4141 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
4201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
4261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
4321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
4381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
4441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
4501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
4561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
4621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
4681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
4741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
4801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
4861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
4921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
4981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
5041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
5101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
5161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
5221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
5281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
5341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
5401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
5461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
5521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
5581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
5641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
5701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
5761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
5821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
5881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
5941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
6001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
6061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
6121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
6181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
6241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
6301 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
6361 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
6421 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
6481 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
6541 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
6601 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
6661 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
6721 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
6781 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
6841 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
6901 tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg
6961 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
7021 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
7081 ccatgtt
//
