| 质粒名称: | pCLE |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 17703 |
| 存储实验室: | Nicholas Gaiano |
|
载体骨架基本信息 |
|
| 载体名称: | pCLE |
| 空载体大小: | 7600 bp |
| 完整质粒大小: | 7702 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: | None |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII(未破坏) |
| 3’克隆位点: | BglII (未破坏) |
| 5’测序引物 | na |
|
文献引用方法 |
|
| 材料和方法部分: | pCLE was a gift from Nicholas Gaiano (Addgene plasmid # 17703 ; http://n2t.net/addgene:17703 ; RRID:Addgene_17703) |
| 参考文献部分: | Radial glial identity is promoted by Notch1 signaling in the murine forebrain. Gaiano N, Nye JS, Fishell G. Neuron. 2000 May . 26(2):395-404. 10.1016/S0896-6273(00)81172-1 PubMed 10839358 |
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 7702 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCLE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7702)
AUTHORS Gaiano N, Nye JS, Fishell G
TITLE Radial glial identity is promoted by Notch1 signaling in the murine
forebrain.
JOURNAL Neuron. 2000 May . 26(2):395-404.
PUBMED 10839358
REFERENCE 2 (bases 1 to 7702)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7702
/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"
misc_feature 2181..2733
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2325..2342)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2552..2571
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2745..4244
/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 4810..5279
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5409..5426)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5580..6168)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5660..5679)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6328..7188)
/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 6951..6970
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7189..7293)
/gene="bla"
/label=AmpR promoter
primer_bind 7361..7379
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7417..7439)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7539..7558
/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 ggtaggaatt ccgcccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
2221 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
2281 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
2341 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
2401 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
2461 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
2521 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
2581 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
2641 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
2701 cgtggttttc ctttgaaaaa cacgatgata atatggggtc gaggctgctg ctgctgctgc
2761 tgctgggcct gaggctacag ctctccctgg gcatcatccc agttgaggag gagaacccgg
2821 acttctggaa ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac
2881 agacagccgc caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga
2941 cagctgccag gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg
3001 ccatggaccg cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc
3061 cagacagtgg agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca
3121 ttggcttgag tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca
3181 tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac
3241 gagtgcagca cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact
3301 cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc
3361 tcatctccaa catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca
3421 tgggaacccc agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg
3481 ggaagaatct ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc
3541 gcactgagct catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg
3601 agcctggaga catgaaatac gagatccacc gagactccac actggacccc tccctgatgg
3661 agatgacaga ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg
3721 tggagggtgg tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg
3781 agacgatcat gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca
3841 cgctgagcct cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc
3901 gagggagctc catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg
3961 tcctcctata cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta
4021 ccgagagcga gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag
4081 agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc
4141 acggcgtgca ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc
4201 cctacaccgc ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccggggc
4261 ggtccgtggt ccccgcgttg cttcctctgc tggccgggac cctgctgctg ctggagacgg
4321 ccactgctcc ctgagtgtcc cgtccctggg gctcctgctt ccccatcccg gagttctcct
4381 gctccccacc tcctgtcgtc ctgcctggcc tccagcccga gtcgtcatcc ccggagtccc
4441 tatacagagg tcctgccatg gaaccttccc ctccccgtgc gctctgggga ctgagcccat
4501 gacaccaaac ctgccccttg gctgctctcg gactccctac cccaacccca gggactgcag
4561 gttgtgccct gtggctgcct gcaccccagg aaaggagggg gctcaggcca tccagccacc
4621 acctacagcc cagtggggtc gacccgataa aataaaagat tttatttagt ctccagaaaa
4681 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
4741 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
4801 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
4861 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
4921 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
4981 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
5041 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
5101 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
5161 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
5221 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
5281 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
5341 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
5401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
5461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
5521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
5581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
5641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
5701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
5761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
5821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
5881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
5941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
6001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
6061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
6121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
6181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
6241 atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag
6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
7321 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
7381 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
7441 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
7501 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
7561 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
7621 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
7681 ttcgctatta cgccagcgcg tt
//
