| 质粒名称: | pCAG-FlpeERT2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14756 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 4779 bp |
| 完整质粒大小: | 7028 bp |
| 载体类型: | 哺乳细胞表达，Flp/FRT |
| 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: | Flpe-ERT2 fusion protein |
| 物种来源: | S. cerevisiae（芽殖酵母） |
| 插入大小: | 2249 bp |
| 基因又名: | inducible Flpe |
| 标签/融合蛋白: | ERT2 (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | pCAG-F |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-FlpeERT2 was a gift from Connie Cepko (爱迪基因 plasmid # 14756) |
| 参考文献部分: | Controlled expression of transgenes introduced by in vivo electroporation. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2007 Jan 5. ():. 10.1073/pnas.0610155104 PubMed 17209010 |
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 pCAG-FlpeERT2 7029 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCAG-FlpeERT2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7029)
AUTHORS Matsuda T, Cepko CL
TITLE Controlled expression of transgenes introduced by in vivo
electroporation.
JOURNAL Proc Natl Acad Sci U S A. 2007 Jan 5. ():.
PUBMED 17209010
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pCAG-FlpeERT2|
FEATURES Location/Qualifiers
source 1..7029
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1678..1697
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1737..3005
/codon_start=1
/product="site-specific recombinase"
/label=FLP
/note="FLP is a site-specific recombinase from
Saccharomyces cerevisiae. Recombination occurs at FRT
sequences."
/translation="MSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI
THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
YPAWNGIISQEVLDYLSSYINRRI"
CDS 3024..3959
/codon_start=1
/product="mutated ligand-binding domain of the human
estrogen receptor (Feil et al., 1997)"
/label=ERT2
/translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
primer_bind complement(4053..4072)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4118..4173
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4172..4191)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4532..4548)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4532..4548)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4545..4567)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4556..4572
/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(4580..4610)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4625..4646
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4705..4900
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4751..4886
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4813..4832
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 4906..5040
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4943..4962)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4997..5016
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5108..5125)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5279..5867)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5359..5378)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6038..6898)
/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 6661..6680
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6899..7003)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgagcc gccaccatga
1741 gccaatttga tatattatgt aaaacaccac ctaaggtcct ggttcgtcag tttgtggaaa
1801 ggtttgaaag accttcaggg gaaaaaatag catcatgtgc tgctgaacta acctatttat
1861 gttggatgat tactcataac ggaacagcaa tcaagagagc cacattcatg agctataata
1921 ctatcataag caattcgctg agtttcgata ttgtcaacaa atcactccag tttaaataca
1981 agacgcaaaa agcaacaatt ctggaagcct cattaaagaa attaattcct gcttgggaat
2041 ttacaattat tccttacaat ggacaaaaac atcaatctga tatcactgat attgtaagta
2101 gtttgcaatt acagttcgaa tcatcggaag aagcagataa gggaaatagc cacagtaaaa
2161 aaatgcttaa agcacttcta agtgagggtg aaagcatctg ggagatcact gagaaaatac
2221 taaattcgtt tgagtatacc tcgagattta caaaaacaaa aactttatac caattcctct
2281 tcctagctac tttcatcaat tgtggaagat tcagcgatat taagaacgtt gatccgaaat
2341 catttaaatt agtccaaaat aagtatctgg gagtaataat ccagtgttta gtgacagaga
2401 caaagacaag cgttagtagg cacatatact tctttagcgc aaggggtagg atcgatccac
2461 ttgtatattt ggatgaattt ttgaggaact ctgaaccagt cctaaaacga gtaaatagga
2521 ccggcaattc ttcaagcaac aaacaggaat accaattatt aaaagataac ttagtcagat
2581 cgtacaacaa ggctttgaag aaaaatgcgc cttatccaat ctttgctata aagaatggcc
2641 caaaatctca cattggaaga catttgatga cctcatttct gtcaatgaag ggcctaacgg
2701 agttgactaa tgttgtggga aattggagcg ataagcgtgc ttctgccgtg gccaggacaa
2761 cgtatactca tcagataaca gcaatacctg atcactactt cgcactagtt tctcggtact
2821 atgcatatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
2881 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
2941 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
3001 gcatatgcgt acgcggatcc tctgctggag acatgagagc tgccaacctt tggccaagcc
3061 cgctcatgat caaacgctct aagaagaaca gcctggcctt gtccctgacg gccgaccaga
3121 tggtcagtgc cttgttggat gctgagcccc ccatactcta ttccgagtat gatcctacca
3181 gacccttcag tgaagcttcg atgatgggct tactgaccaa cctggcagac agggagctgg
3241 ttcacatgat caactgggcg aagagggtgc caggctttgt ggatttgacc ctccatgatc
3301 aggtccacct tctagaatgt gcctggctag agatcctgat gattggtctc gtctggcgct
3361 ccatggagca cccagtgaag ctactgtttg ctcctaactt gctcttggac aggaaccagg
3421 gaaaatgtgt agagggcatg gtggagatct tcgacatgct gctggctaca tcatctcggt
3481 tccgcatgat gaatctgcag ggagaggagt ttgtgtgcct caaatctatt attttgctta
3541 attctggagt gtacacattt ctgtccagca ccctgaagtc tctggaagag aaggaccata
3601 tccaccgagt cctggacaag atcacagaca ctttgatcca cctgatggcc aaggcaggcc
3661 tgaccctgca gcagcagcac cagcggctgg cccagctcct cctcatcctc tcccacatca
3721 ggcacatgag taacaaaggc atggagcatc tgtacagcat gaagtgcaag aacgtggtgc
3781 ccctctatga cctgctgctg gaggcggcgg acgcccaccg cctacatgcg cccactagcc
3841 gtggaggggc atccgtggag gagacggacc aaagccactt ggccactgcg ggctctactt
3901 catcgcattc cttgcaaaag tattacatca cgggggaggc agagggtttc cctgccacag
3961 cttgatagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
4021 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
4081 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
4141 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
4201 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg
4261 ctgccatgaa caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
4321 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
4381 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
4441 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct
4501 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
4561 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
4621 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
4681 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
4741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
4801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
4861 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
4921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
4981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
5041 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
5101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
5161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
5221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
5281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
5341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
5401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
5461 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
5521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
5581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
5641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
5701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
5761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
5821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
5881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
5941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
6001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
6061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
6121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
6181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
6241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
6301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
6361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
6421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
6481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
6541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
6601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
6661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
6721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
6781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
6841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
7021 tgccacctg
//
