| 质粒名称: | TRE-mCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 92202 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | AAV vector |
| 空载体大小: | 4037 bp |
| 完整质粒大小: | 5119 bp |
| 载体类型: | 哺乳细胞表达,AAV |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 物种来源: | Synthetic |
| 插入大小: | 738 bp |
| 启动子: | TRE |
| 克隆方法: | Gibson 克隆 |
| 5’测序引物 | gcggccgcACGCGTccttc |
| 3’测序引物 | CATAGTTAAGAATACCAGTCAATCTTTCAC |
|
文献引用方法 |
|
| 材料和方法部分: | TRE-mCherry was a gift from Alice Ting (Addgene plasmid # 92202 ; http://n2t.net/addgene:92202 ; RRID:Addgene_92202) |
| 参考文献部分: | A light- and calcium-gated transcription factor for imaging and manipulating activated neurons. Wang W, Wildes CP, Pattarabanjird T, Sanchez MI, Glober GF, Matthews GA, Tye KM, Ting AY. Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909. 10.1038/nbt.3909 PubMed 28650461 |
买家导航
TREmCherry载体基本信息
TREmCherry载体图谱质粒图谱和多克隆位点信息
TREmCherry载体简介
TREmCherry载体序列
LOCUS TRE-mCherry 5119 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Fluorescent protein reporter construct.
ACCESSION .
VERSION .
KEYWORDS TRE-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5119)
AUTHORS Wang W, Wildes CP, Pattarabanjird T, Sanchez MI, Glober GF, Matthews
GA, Tye KM, Ting AY
TITLE A light- and calcium-gated transcription factor for imaging and
manipulating activated neurons.
JOURNAL Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909.
PUBMED 28650461
REFERENCE 2 (bases 1 to 5119)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5119
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 8..137
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
protein_bind 183..201
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 218..236
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 254..272
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 290..308
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 325..343
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 361..379
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 391..429
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 426..450
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 478..487
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 505..514
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 511..1221
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(656..674)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 924..943
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 1253..1841
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1306..1326)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1724..1735)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 1873..2349
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2072..2091)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 2389..2529
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 2389..2518
/label=AAV2 ITR
rep_origin 2604..3059
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2691..2710)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2901..2922
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3076..3095)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3195..3217
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3255..3273)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3341..3445
/gene="bla"
/label=AmpR promoter
CDS 3446..4306
/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 complement(3664..3683)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4477..5065
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4966..4985
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
61 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
121 catcactagg ggttcctgcg gccgcacgcg tccttcaggt ccgaggttct agacgagttt
181 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
241 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
301 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
361 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata
421 agcagagctc gtttagtgaa ccgtcagatc gcaaagggcg aattcgatcc accggtcgcc
481 accatggctc gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg
541 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
601 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
661 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
721 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
781 tccttccccg agggattcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
841 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
901 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
961 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
1021 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
1081 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
1141 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
1201 atggacgagc tgtacaagta aactagtacc ggtagatatc aagcttatcg ataatcaacc
1261 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
1321 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
1381 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
1441 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
1501 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
1561 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
1621 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
1681 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
1741 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
1801 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
1861 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
1921 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
1981 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
2041 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
2101 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
2161 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
2221 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
2281 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
2341 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
2401 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
2461 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
2521 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
2581 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
2641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
2701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
2761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
2821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
2881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
2941 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
3001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
3241 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
3301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
3421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
3481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
3541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
3601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
3661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
3721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
3781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
3841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
3901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
3961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
4021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
4081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
4141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
4201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
4261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
4321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
5101 cttttgctgg ccttttgct
//
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TREmCherry载体质粒应用举例
pAAV-tre-mcherry
2020-02-20
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