| 质粒名称: | Sema3e(S)-Fc-His |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 72146 |
| 存储实验室: | Woj Wojtowicz |
|
载体骨架基本信息 |
|
| 载体名称: | pCMVi-SV40ori |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达 |
| 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: | Sema3e,Semaphorin 3E,NCBI gene ID 20349; Reference sequence NM_011348.2 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1662 bp |
| 启动子 | CMV |
| 标签/融合蛋白: | Fc His（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NotI(未破坏) |
| 3' 克隆位点: | AscI(未破坏) |
| 5' 测序引物 | GTCAGAGGTAACTCCCGTTGC |
| 3' 测序引物 | ATCATGAGGGTGTCCTTG |
|
文献引用方法 |
|
| 材料和方法部分: | Sema3e(S)-Fc-His was a gift from Woj Wojtowicz (Addgene plasmid # 72146 ; http://n2t.net/addgene:72146 ; RRID:Addgene_72146) |
| 参考文献部分: | An extracellular biochemical screen reveals that FLRTs and Unc5s mediate neuronal subtype recognition in the retina. Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray TA, Kay JN, Wojtowicz WM. Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149. 10.7554/eLife.08149 PubMed 26633812 |
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
-Fc-His质粒图谱.png)
载体简介
载体序列
LOCUS Sema3e(S)-Fc-His 7388 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses the Sema3E protein (truncated at cleavage site P1; ie,
short), C-terminally fused to the Fc region of human IgG1 + 6X
histidine tag..
ACCESSION .
VERSION .
KEYWORDS Sema3e(S)-Fc-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7388)
AUTHORS Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray
TA, Kay JN, Wojtowicz WM
TITLE An extracellular biochemical screen reveals that FLRTs and Unc5s
mediate neuronal subtype recognition in the retina.
JOURNAL Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
PUBMED 26633812
REFERENCE 2 (bases 1 to 7388)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7388
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 105..408
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 409..612
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 563..583
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 609..633
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 748..1573
/label=CMV intron A
/note="human cytomegalovirus intron A (Chapman et al.,
1991)"
regulatory 1604..1613
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3968..3985
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 4011..4132
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4048..4067)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4102..4121
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4170..4187)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4341..4929)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4421..4440)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5100..5960)
/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 5723..5742
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5961..6065)
/gene="bla"
/label=AmpR promoter
primer_bind 6133..6151
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6189..6211)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6213..6233)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6218..6548
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6399..6534
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6461..6480
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 6685..6704
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(6771..7226)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6908..6929)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7120..7139
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7352..7374
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7366..7383
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7367..7383
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tggcaccggc
1621 cggacacatc ctcaccttgc tgctctgggg tcacctgctg gaactctgga ccccaggtca
1681 ctccgcgaac ccctcctacc ccaggctacg cctgtcacat aaagaacttt tggaactgaa
1741 taggacttca atatttcaaa gcccccttgg atttcttgat ctccatacaa tgctgctgga
1801 tgagtatcaa gaacggctct ttgtgggagg cagagacctt gtctattccc tgaacttgga
1861 acgagtcagt gacggctaca gagagatata ctggccgagc acagcagtaa aggtagaaga
1921 atgcataatg aaaggaaaag acgcaaatga gtgtgccaat tatatccggg ttttgcatca
1981 ctacaacagg acacaccttc tgacctgtgc tactggagct tttgatccac actgtgcctt
2041 catcagagtc gggcaccatt cagaggaacc cctgtttcac ctggagtcac acagatctga
2101 gagaggaagg ggcagatgtc cttttgaccc caactcctcc tttgtgtcca cgctagttgg
2161 gaatgagctg tttgctggac tctacagtga ctattggggc agagactcgg cgatcttccg
2221 cagcatgggg aagttaggcc atattcgcac tgagcatgac gatgagcggc tcctgaaaga
2281 accaaaattt gtaggttcat atatgattcc tgataacgaa gaccgagatg acaacaaaat
2341 gtactttttc tttactgaga aggcgctgga ggcggagaac aacgcccaca cgatctacac
2401 ccgagtgggg cggctgtgcg tgaatgacat gggaggacag agaatcctgg tgaacaagtg
2461 gagcactttc cttaaagcgc ggctggtttg ctcagtgccg ggaatgaatg gaatcgacac
2521 atactttgac gaactagagg atgtgttttt actgccgacc agagatccta agaatccagt
2581 gatatttgga ctgtttaata ctaccagcaa tatatttaga ggccatgctg tatgtgtgta
2641 tcacatgtca agtatccggg aagcctttaa tggcccatat gctcataaag aaggccctga
2701 ataccactgg tcactatatg aaggaaaagt cccctaccca aggcctggtt cctgtgccag
2761 caaagtaaac ggaggcaagt atggaaccac caaagattac cccgatgacg ccatccggtt
2821 cgcaaggatg catcctctaa tgtatcagcc cataaaacct gttcataaaa aaccaatact
2881 ggtaaaaaca gatggaaaat acaacctgag gcaacttgcc gtggatcggg tggaagcgga
2941 ggatggccag tatgacgtct tatttattgg gacagacaca ggaattgtgc tgaaagtaat
3001 cacaatttac aaccaagaaa cagagtggat ggaggaagtc attctagagg aacttcaaat
3061 attcaaggat ccagccccta tcatttctat ggaaatttct tcaaagagac aacagcttta
3121 cattggatca gcctctgctg tggcacaagt cagattccat cactgcgaca tgtatggcag
3181 tgcttgtgct gactgctgcc tggctcgaga cccgtactgt gcctgggatg gcatatcctg
3241 ctccaggtac tacccaacag gtgcacacgc aggcgcgcct gacaaaactc acacatgccc
3301 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
3361 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
3421 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
3481 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
3541 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
3601 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
3661 ggtgtatacc ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg
3721 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
3781 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
3841 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
3901 gatgcatgag gctctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
3961 aggcggccat catcaccatc accattaatc tagagatctt attaaagcag aacttgttta
4021 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
4081 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
4141 ggtcgactct agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4201 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4261 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4321 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4381 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4441 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4501 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4561 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4621 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4681 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4741 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
4801 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4861 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4921 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4981 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
5041 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
5101 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
5161 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
5221 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
5281 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
5341 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
5401 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
5461 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
5521 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
5581 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
5641 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
5701 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5761 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5821 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5881 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
5941 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
6001 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
6061 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
6121 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
6181 ggtgaaaacc tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa
6241 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
6301 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
6361 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
6421 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
6481 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
6541 tttgcaaaaa gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt
6601 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
6661 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
6721 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
6781 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
6841 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
6901 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
6961 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
7021 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
7081 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
7141 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
7201 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
7261 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
7321 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
7381 agtgaatt
//
