| 质粒名称: | pCLE NICD3 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 26894 |
| 存储实验室: | Nicholas Gaiano |
|
载体骨架基本信息 |
|
| 载体名称: | pCLE |
| 完整质粒大小: | 9716 bp |
| 载体类型: | 哺乳细胞表达、Retroviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | NICD3 / Notch3 intracellular domain |
| 物种来源: | H. sapiens (human) |
| 标签/融合蛋白: |
IRES (C terminal on backbone)
Alkaline Phosphatase (C terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | EF-1a-F |
|
文献引用方法 |
|
| 材料和方法部分: | pCLE NICD3 was a gift from Nicholas Gaiano (Addgene plasmid # 26894 ; http://n2t.net/addgene:26894 ; RRID:Addgene_26894) |
| 参考文献部分: | Notch3 signaling promotes radial glial/progenitor character in the mammalian telencephalon. Dang L, Yoon K, Wang M, Gaiano N. Dev Neurosci. 2006;28(1-2):58-69. 10.1159/000090753 PubMed 16508304 |
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 pCLE NICD3 9716 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCLE NICD3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9716)
AUTHORS Dang L, Yoon K, Wang M, Gaiano N
TITLE Notch3 signaling promotes radial glial/progenitor character in the
mammalian telencephalon.
JOURNAL Dev Neurosci. 2006;28(1-2):58-69.
PUBMED 16508304
REFERENCE 2 (bases 1 to 9716)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9716
/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"
misc_feature 823..1022
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1223..1639
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1569..1591
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1607..1623
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 2167..2176
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 4195..4747
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4339..4356)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4566..4585
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4759..6258
/codon_start=1
/product="secreted alkaline phosphatase from human
placenta"
/label=SEAP
/note="The natural enzyme has a C-terminal transmembrane
sequence that is replaced by a GPI anchor."
/translation="LLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPA
QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHL
MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRA
YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKAR
DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
LTR 6824..7293
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(7423..7440)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7594..8182)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7674..7693)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8342..9202)
/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 8965..8984
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9203..9307)
/gene="bla"
/label=AmpR promoter
primer_bind 9375..9393
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(9431..9453)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 9553..9572
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
601 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
661 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
721 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
781 ggagacccct gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta
841 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
1621 cactccttct ctaggcgccg gccggatcga gcagggggat catctaatca agcacaaata
1681 aggggcgtgt aacacaaaag ccagcgaccc tttccaatgc aaatcaaact tgcaattctt
1741 tgccgttttt atcatttaag tgtcggctta aggtccacta tcagatgtaa acagccttat
1801 ctaacaaagg tatcattaca ttctgaaatt ctcaggcatg caagctagct tatgacgcac
1861 tagggagtgc cacccttcct ttcgccctaa cttcgtgata actcgcgcgt ttcactcaac
1921 agctgcatcc gccctagtgc tactgggagt tgtagtatac aagacgctta caggctgaat
1981 gttctgtcaa gaccccgcct ctagcacttt gggaattctg gacttgatga tgtcatggtt
2041 aatccccgcc cagtagaggc ggctatataa agggtggtta aggcccggtt cgctctcttc
2101 ctcaccgggt ctgcggcgag ttctagctga agctgatccc agtgtggtgg taccagatct
2161 cgaccaccat ggtggcccgg cgcaagcgcg agcacagcac cctctggttc cctgagggct
2221 tctcactgca caaggacgtg gcctctggtc acaagggccg gcgggaaccc gtgggccagg
2281 acgcgctggg catgaagaac atggccaagg gtgagagcct gatgggggag gtggccacag
2341 actggatgga cacagagtgc ccagaggcca agcggctaaa ggtagaggag ccaggcatgg
2401 gggctgagga ggctgtggat tgccgtcagt ggactcaaca ccatctggtt actgctgaca
2461 tccgcgtggc accagccatg gcactgacac caccacaggg cgacgcagat gctgatggca
2521 tggatgtcaa tgtgcgtggc ccagatggct tcaccccgct aatgctggct tccttctgtg
2581 ggggggctct ggagccaatg ccaactgaag aggatgaggc agatgacaca tcagctagca
2641 tcatctccga cctgatctgc cagggggctc agcttggggc acggactgac cgtactggcg
2701 agactgcttt gcacctggct gcccgttatg cccgtgctga tgcagccaag cggctgctgg
2761 atgctggggc agacaccaat gcccaggacc actcaggccg cactcccctg cacacagctg
2821 tcacagccga tgcccagggt gtcttccaga ttctcatccg aaaccgctct acagacttgg
2881 atgcccgcat ggcagatggc tcaacggcac tgatcctggc ggcccgcctg gcagtagagg
2941 gcatggtgga agagctcatc gccagccatg ctgatgtcaa tgctgtggat gagcttggga
3001 aatcagcctt acactgggct gcggctgtga acaacgtgga agccactttg gccctgctca
3061 aaaatggagc caataaggac atgcaggata gcaaggagga gaccccccta ttcctggccg
3121 cccgcgaggg cagctatgag gctgccaagc tgctgttgga ccactttgcc aaccgtgaga
3181 tcaccgacca cctggacagg ctgccgcggg acgtagccca ggagagactg caccaggaca
3241 tcgtgcgctt gctggatcaa cccagtgggc cccgcagccc ccccggtccc cacggcctgg
3301 ggcctctgct ctgtcctcca ggggccttcc tccctggcct caaagcggca cagtcggggt
3361 ccaagaagag caggaggccc cccgggaagg cggggctggg gccgcagggg ccccgggggc
3421 ggggcaagaa gctgacgctg gcctgcccgg gccccctggc tgacagctcg gtcacgctgt
3481 cgcccgtgga ctcgctggac tccccgcggc ctttcggtgg gccccctgct tcccctggtg
3541 gcttccccct tgaggggccc tatgcagctg ccactgccac tgcagtgtct ctggcacagc
3601 ttggtggccc aggccgggcg ggtctagggc gccagccccc tggaggatgt gtactcagcc
3661 tgggcctgct gaaccctgtg gctgtgcccc tcgattgggc ccggctgccc ccacctgccc
3721 ctccaggccc ctcgttcctg ctgccactgg cgccgggacc ccagctgctc aacccaggga
3781 cccccgtctc cccgcaggag cggcccccgc cttacctggc agtcccagga catggcgagg
3841 agtacccggc ggctggggca cacagcagcc ccccaaaggc ccgcttcctg cgggttccca
3901 gtgagcaccc ttacctgacc ccatcccccg aatcccctga gcactgggcc agcccctcac
3961 ctccctccct ctcagactgg tccgaatcca cgcctagccc agccactgcc actggggcca
4021 tggccaccac cactggggca ctgcctgccc agccacttcc cttgtctgtt cccagctccc
4081 ttgctcaggc ccagacccag ctggggcccc agccggaagt tacccccaag aggcaagtgt
4141 tggcctgaga cgctcgtcag ttcttagatc gatctggtag gaattccccc cctaacgtta
4201 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
4261 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
4321 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
4381 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
4441 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
4501 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
4561 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
4621 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
4681 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
4741 gataatatgg ggtcgaggct gctgctgctg ctgctgctgg gcctgaggct acagctctcc
4801 ctgggcatca tcccagttga ggaggagaac ccggacttct ggaaccgcga ggcagccgag
4861 gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa cctcatcatc
4921 ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct aaaagggcag
4981 aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc atatgtggct
5041 ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac agccacggcc
5101 tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc cgcccgcttt
5161 aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa
5221 gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc gccagccggc
5281 acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc
5341 cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga cattgacgtg
5401 atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc tgagtaccca
5461 gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca ggaatggctg
5521 gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca ggcttccctg
5581 gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa atacgagatc
5641 caccgagact ccacactgga cccctccctg atggagatga cagaggctgc cctgcgcctg
5701 ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt
5761 catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga cgacgccatt
5821 gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac tgccgaccac
5881 tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt cgggctggcc
5941 cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa cggtccaggc
6001 tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg gagccccgag
6061 tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga ggacgtggcg
6121 gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca gaccttcata
6181 gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga cctggcgccc
6241 cccgccggca ccaccgacgc cgcgcacccg gggcggtccg tggtccccgc gttgcttcct
6301 ctgctggccg ggaccctgct gctgctggag acggccactg ctccctgagt gtcccgtccc
6361 tggggctcct gcttccccat cccggagttc tcctgctccc cacctcctgt cgtcctgcct
6421 ggcctccagc ccgagtcgtc atccccggag tccctataca gaggtcctgc catggaacct
6481 tcccctcccc gtgcgctctg gggactgagc ccatgacacc aaacctgccc cttggctgct
6541 ctcggactcc ctaccccaac cccagggact gcaggttgtg ccctgtggct gcctgcaccc
6601 caggaaagga gggggctcag gccatccagc caccacctac agcccagtgg ggtcgacccg
6661 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
6721 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
6781 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
6841 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
6901 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
6961 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
7021 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
7081 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
7141 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
7201 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
7261 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
7321 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
7381 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
7441 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
7501 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
7561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
7621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
7681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
7801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
7861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
7921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
7981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
8041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
8101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
8161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
8221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
8281 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
8341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
8401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
8461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
8521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
8581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
8641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
8701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
8761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
8821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
8881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
8941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
9001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
9061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
9121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
9181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
9241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
9301 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
9361 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
9421 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
9481 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
9541 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
9601 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
9661 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cgcgtt
//
