| 质粒名称: | pCAG-ERT2CreERT2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13777 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4779 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,Cre/Lox |
| 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: | ERT2-Cre-ERT2 fusion protein,inducible Cre |
| 物种来源: | Bacteriophage P1 |
| 插入大小: | 2951 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: | EcoRI(未破坏) |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | pCAG-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-ERT2CreERT2 was a gift from Connie Cepko (Addgene plasmid # 13777 ; http://n2t.net/addgene:13777 ; RRID:Addgene_13777) |
| 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
pCAGERT2CreERT2载体基本信息
pCAGERT2CreERT2载体图谱质粒图谱和多克隆位点信息
pCAGERT2CreERT2载体简介
pCAGERT2CreERT2载体序列
LOCUS pCAG-ERT2CreERT2 7740 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION Mammalian expression of conditionally active form of Cre recombinase
(activated in response to tamoxifen).
ACCESSION .
VERSION .
KEYWORDS pCAG-ERT2CreERT2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7740)
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
REFERENCE 2 (bases 1 to 7740)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7740
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1686..1705
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1744..1753
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1753..2688
/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"
CDS 2698..3720
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
D"
CDS 3733..4668
/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(4762..4781)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4827..4882
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4881..4900)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5243..5259)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5243..5259)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5256..5278)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5267..5283
/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(5291..5321)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5336..5357
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5416..5611
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5462..5597
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5524..5543
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5617..5751
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5654..5673)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5708..5727
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5819..5836)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5990..6578)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6070..6089)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6749..7609)
/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 7372..7391
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7610..7714)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggsgcgc gccrgggggg gggggggggg gggggggggg
541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcccgggtga
1741 gccgccacca tggctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc
1801 aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc
1861 ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt
1921 gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc
1981 aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt
2041 ctagaatgtg cctggctaga gatcctgatg attggtctcg tctggcgctc catggagcac
2101 ccagtgaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta
2161 gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg
2221 aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg
2281 tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc
2341 ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag
2401 cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt
2461 aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac
2521 ctgctgctgg aggcggcgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca
2581 tccgtggagg agacggacca aagccacttg gccactgcgg gctctacttc atcgcattcc
2641 ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagc tgtcgacaat
2701 ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt
2761 cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca tacctggaaa
2821 atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg
2881 tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc gcgcggtctg
2941 gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg tcggtccggg
3001 ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat ccgaaaagaa
3061 aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac
3121 caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa tctggcattt
3181 ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat cagggttaaa
3241 gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac gaaaacgctg
3301 gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact ggtcgagcga
3361 tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg ccgggtcaga
3421 aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc cctggaaggg
3481 atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg tcagagatac
3541 ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc ccgcgctgga
3601 gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat tgtcatgaac
3661 tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga agatggcgat
3721 ctcgagccat ctgctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc
3781 aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc
3841 ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt
3901 gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc
3961 aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt
4021 ctagaatgtg cctggctaga gatcctgatg attggtctcg tctggcgctc catggagcac
4081 ccagtgaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta
4141 gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg
4201 aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg
4261 tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc
4321 ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag
4381 cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt
4441 aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac
4501 ctgctgctgg aggcggcgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca
4561 tccgtggagg agacggacca aagccacttg gccactgcgg gctctacttc atcgcattcc
4621 ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagc ttgatagcgg
4681 ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
4741 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
4801 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
4861 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
4921 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
4981 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
5041 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
5101 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
5161 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
5221 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
5281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
5341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
5401 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
5461 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
5521 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
5581 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
5641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
5701 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
6541 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
6601 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
6661 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
6721 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
6781 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
6841 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
6901 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
6961 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
7021 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
7081 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
7141 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
7201 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
7261 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
7321 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
7381 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
7441 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
7501 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
7561 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
7621 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
7681 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
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