| 质粒名称: | pEF-hSEMA4C-Fc |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74449 |
| 存储实验室: | Roland Friedel |
|
载体骨架基本信息 |
|
| 载体名称: | pEF-BOS |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | 8696 bp |
| 载体修饰: | 质粒pEF-BOS-hSEMA4D-Fc（Kumanogoh等人，2000）通过切断Sema4D外域并插入SEMA4C外域片段进行修饰。 |
| 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: | human Semaphorin-4C ectodomain with human Fc tag |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3261 bp |
| 基因突变: | -- |
| GenBank编号 | NM_017789.4 |
| 启动子 | EF1-alpha |
| 标签/融合蛋白: | human Fc tag（插入C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | EF-F CATTCTCAAGCCTCAGACAGTGGT |
|
文献引用方法 |
|
| 材料和方法部分: | pEF-hSEMA4C-Fc was a gift from Roland Friedel (Addgene plasmid # 74449 ; http://n2t.net/addgene:74449 ; RRID:Addgene_74449) |
| 参考文献部分: | Plexin-B2 promotes invasive growth of malignant glioma. Le AP, Huang Y, Pingle SC, Kesari S, Wang H, Yong RL, Zou H, Friedel RH. Oncotarget. 2015 Mar 30;6(9):7293-304. 10.18632/oncotarget.3421 PubMed 25762646 |
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 pEF-hSEMA4C-Fc 8696 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses human Semaphorin-4C ectodomain fused to human Fc in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pEF-hSEMA4C-Fc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8696)
AUTHORS Le AP, Huang Y, Pingle SC, Kesari S, Wang H, Yong RL, Zou H, Friedel
RH
TITLE Plexin-B2 promotes invasive growth of malignant glioma.
JOURNAL Oncotarget. 2015 Mar 30;6(9):7293-304.
PUBMED 25762646
REFERENCE 2 (bases 1 to 8696)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8696
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 96..200
/gene="bla"
/label=AmpR promoter
CDS 201..1061
/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(419..438)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1232..1820
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1721..1740
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1974..1991
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2108..2129
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2144..2174
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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)."
primer_bind 2187..2209
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2206..2222
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 2251..2461
/label=SV40 promoter
/note="SV40 early promoter"
promoter 2273..2469
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 2320..2455
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2382..2401
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2558..3736
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2789..3727
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3684..3704
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind complement(7826..7843)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7826..7842)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7835..7857)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(8110..8490)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8172..8193)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 8384..8403
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(8527..8546)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8646..8668
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
2221 accatgatta cgccaagctt aggctcccca gcaggcagaa gtatgcaaag catgcatctc
2281 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
2341 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
2401 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
2461 ttttgcaaaa agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
2521 aatggacctt ctaggtcttg aaaggagtgc ctcgtgaggc tccggtgccc gtcagtgggc
2581 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
2641 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
2701 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
2761 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
2821 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
2881 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
2941 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
3001 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
3061 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
3121 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
3181 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
3241 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc
3301 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
3361 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
3421 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
3481 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
3541 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
3601 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
3661 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
3721 atttcaggtg tcgtgaggaa ttctctagag atccctcgac ctcgagatcc atcgggcccc
3781 ccctcgaggt cgactcgcct ctacctgatg gccccacact gggctgtctg gctgctggca
3841 gcaaggctgt ggggcctggg cattggggct gaggtgtggt ggaaccttgt gccgcgtaag
3901 acagtgtctt ctggggagct ggccacggta gtacggcggt tctcccagac cggcatccag
3961 gacttcctga cactgacgct gacggagccc actgggcttc tgtacgtggg cgcccgagag
4021 gccctgtttg ccttcagcat ggaggccctg gagctgcaag gagcgatctc ctgggaggcc
4081 cccgtggaga agaagactga gtgtatccag aaagggaaga acaaccagac cgagtgcttc
4141 aacttcatcc gcttcctgca gccctacaat gcctcccacc tgtacgtctg tggcacctac
4201 gccttccagc ccaagtgcac ctacgtcaac atgctcacct tcactttgga gcatggagag
4261 tttgaagatg ggaagggcaa gtgtccctat gacccagcta agggccatgc tggccttctt
4321 gtggatggtg agctgtactc ggccacactc aacaacttcc tgggcacgga acccattatc
4381 ctgcgtaaca tggggcccca ccactccatg aagacagagt acctggcctt ttggctcaac
4441 gaacctcact ttgtaggctc tgcctatgta cctgagagtg tgggcagctt cacgggggac
4501 gacgacaagg tctacttctt cttcagggag cgggcagtgg agtccgactg ctatgccgag
4561 caggtggtgg ctcgtgtggc ccgtgtctgc aagggcgata tggggggcgc acggaccctg
4621 cagaggaagt ggaccacgtt cctgaaggcg cggctggcat gctctgcccc gaactggcag
4681 ctctacttca accagctgca ggcgatgcac accctgcagg acacctcctg gcacaacacc
4741 accttctttg gggtttttca agcacagtgg ggtgacatgt acctgtcggc catctgtgag
4801 taccagttgg aagagatcca gcgggtgttt gagggcccct ataaggagta ccatgaggaa
4861 gcccagaagt gggaccgcta cactgaccct gtacccagcc ctcggcctgg ctcgtgcatt
4921 aacaactggc atcggcgcca cggctacacc agctccctgg agctacccga caacatcctc
4981 aacttcgtca agaagcaccc gctgatggag gagcaggtgg ggcctcggtg gagccgcccc
5041 ctgctcgtga agaagggcac caacttcacc cacctggtgg ccgaccgggt tacaggactt
5101 gatggagcca cctatacagt gctgttcatt ggcacaggag acggctggct gctcaaggct
5161 gtgagcctgg ggccctgggt tcacctgatt gaggagctgc agctgtttga ccaggagccc
5221 atgagaagcc tggtgctatc tcagagcaag aagctgctct ttgccggctc ccgctctcag
5281 ctggtgcagc tgcccgtggc cgactgcatg aagtatcgct cctgtgcaga ctgtgtcctc
5341 gcccgggacc cctattgcgc ctggagcgtc aacaccagcc gctgtgtggc cgtgggtggc
5401 cactctggat ctctactgat ccagcatgtg atgacctcgg acacttcagg catctgcaac
5461 ctccgtggca gtaagaaagt caggcccact cccaaaaaca tcacggtggt ggcgggcaca
5521 gacctggtgc tgccctgcca cctctcctcc aacttggccc atgcccgctg gacctttggg
5581 ggccgggacc tgcctgcgga acagcccggg tccttcctct acgatgcccg gctccaggcc
5641 ctggttgtga tggctgccca gccccgccat gccggggcct accactgctt ttcagaggag
5701 cagggggcgc ggctggctgc tgaaggctac cttgtggctg tcgtggcagg cccgtcggtg
5761 accttggagg cccgggcccc cctggaaaac caaagtaagt aggatccact agttctatcc
5821 tgcctggacg catcccggct atgcagtccc agtccagggc agcaaggcag gccccgtctg
5881 cctcttcacc cggaggcctc tgcccgcccc actcatgctc agggagaggg tcttctggct
5941 ttttccccag gctctgggca ggcacaggct aggtgcccct aacccaggcc ctgcacacaa
6001 aggggcaggt gctgggctca gacctgccaa gagccatatc cgggaggacc ctgcccctga
6061 cctaagccca ccccaaaggc caaactctcc actccctcag ctcggacacc ttctctcctc
6121 ccagattcca gtaactccca atcttctctc tgcagagccc aaatcttgtg acaaaactca
6181 cacatgccca ccgtgcccag gtaagccagc ccaggcctcg ccctccagct caaggcggga
6241 caggtgccct agagtagcct gcatccaggg acaggcccca gccgggtgct gacacgtcca
6301 cctccatctc ttcctcagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc
6361 caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg
6421 acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc
6481 ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg
6541 tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca
6601 acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggt gggacccgtg
6661 gggtgcgagg gccacatgga cagaggccgg ctcggcccac cctctgccct gagagtgacc
6721 gctgtaccaa cctctgtccc tacagggcag ccccgagaac cacaggtgta caccctgccc
6781 ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt caaaggcttc
6841 tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag
6901 accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctatagcaa gctcaccgtg
6961 gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg
7021 cacaaccact acacgcagaa gagcctctcc ctgtccccgg gtaaatgagt gcgacggccg
7081 gcaagccccc gctccccggg ctctcgcggt cgcacgagga tgcttggcac gtaccccgtc
7141 tacatacttc ccaggcaccc agcatggaaa taaagcaccc accactgccc tagagcggcc
7201 gcgactctag agtgagggtc cccacctggg acccttgaga gtatcaggtc tcccacgtgg
7261 gagacaagaa atccctgttt aatatttaaa cagcagtgtt ccccatctgg gtccttgcac
7321 ccctcactct ggcctcagcc gactgcacag cggcccctgc atccccttgg ctgtgaggcc
7381 cctggacaag cagaggtggc cagagctggg aggcatggcc ctggggtccc acgaatttgc
7441 tggggaatct cgtttttctt cttaagactt ttgggacatg gtttgactcc cgaacatcac
7501 cgacgcgtct cctgtttttc tgggtggcct cgggacacct gccctgcccc cacgagggtc
7561 aggactgtga ctctttttag ggccaggcag gtgcctggac atttgccttg ctggacgggg
7621 actggggatg tgggagggag cagacaggag gaatcatgtc aggcctgtgt gtgaaaggaa
7681 gctccactgt caccctccac ctcttcaccc cccactcacc agtgtcccct ccactgtcac
7741 attgtaactg aacttcagga taataaagtg cttgcctcca aaaaaaaaaa aaaaaaaaaa
7801 aaaaaaaaaa aaaaaaaaag aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
7861 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
7921 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
7981 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaaaacgt
8041 taatattttg ttaaaattcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
8101 atggtggttc cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt
8161 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
8221 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
8281 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
8341 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
8401 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
8461 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgag catggtgcac
8521 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
8581 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
8641 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
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