| 质粒名称: | pcDNA3-OVA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64599 |
| 存储实验室: | Sandra Diebold, Martin Zenke |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6718 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Ovalbumin |
| 物种来源: | G. gallus (chicken) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | CMV Forward |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-OVA was a gift from Sandra Diebold & Martin Zenke (Addgene plasmid # 64599 ; http://n2t.net/addgene:64599 ; RRID:Addgene_64599) |
| 参考文献部分: | MHC class II presentation of endogenously expressed antigens by transfected dendritic cells. Diebold SS, Cotten M, Koch N, Zenke M. Gene Ther. 2001 Mar;8(6):487-93. 10.1038/sj.gt.3301433 PubMed 11313828 |
买家导航
pcDNA3OVA载体基本信息
pcDNA3OVA载体图谱质粒图谱和多克隆位点信息
pcDNA3OVA载体简介
pcDNA3OVA载体序列
LOCUS pcDNA3-OVA 6711 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expression of chicken ovalbumin as model antigen for study of MHC
class I and class II presentation.
ACCESSION .
VERSION .
KEYWORDS pcDNA3-OVA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6711)
AUTHORS Diebold SS, Cotten M, Koch N, Zenke M
TITLE MHC class II presentation of endogenously expressed antigens by
transfected dendritic cells.
JOURNAL Gene Ther. 2001 Mar;8(6):487-93.
PUBMED 11313828
REFERENCE 2 (bases 1 to 6711)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6711
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(85..105)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 90..419
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 270..405
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 332..351
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 486..1280
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(540..559)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1150..1169
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 1454..1575
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1491..1510)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1545..1564
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1624..1640)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1624..1640)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1637..1659)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1648..1664
/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)."
promoter complement(1672..1702)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1717..1738
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1855..1872)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2026..2614)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2106..2125)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2785..3645)
/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 3408..3427
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3646..3750)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3825..3844)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4016..4395
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4396..4599
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4550..4570
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 4644..4663
/label=T7
/note="T7 promoter, forward primer"
promoter 4644..4662
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4750..5910
/codon_start=1
/product="chicken ovalbumin"
/label=ovalbumin
/translation="MGSIGAASMEFCFDVFKELKVHHANENIFYCPIAIMSALAMVYLG
AKDSTRTQINKVVRFDKLPGFGDSIEAQCGTSVNVHSSLRDILNQITKPNDVYSFSLAS
RLYAEERYPILPEYLQCVKELYRGGLEPINFQTAADQARELINSWVESQTNGIIRNVLQ
PSSVDSQTAMVLVNAIVFKGLWEKAFKDEDTQAMPFRVTEQESKPVQMMYQIGLFRVAS
MASEKMKILELPFASGTMSMLVLLPDEVSGLEQLESIINFEKLTEWTSSNVMEERKIKV
YLPRMKMEEKYNLTSVLMAMGITDVFSSSANLSGISSAESLKISQAVHAAHAEINEAGR
EVVGSAEAGVDAASVSEEFRADHPFLFCIKHIATNAVLFFGRCVSP"
promoter complement(6045..6063)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6046..6063)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(6083..6100)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6089..6313
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6359..76
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6446..6465)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6656..6677
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
61 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
421 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
481 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
541 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
601 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
661 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
721 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
781 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
841 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
901 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
961 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
1021 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
1081 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
1141 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
1201 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
1261 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
1321 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
1381 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
1441 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
1501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
1561 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
1621 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
1681 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
1741 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
1801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
1861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
1921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
1981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
2041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
2101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
2161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
2221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
2281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
2341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
2401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
2461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
2521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
2581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
2641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
2701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
2761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
2821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
2881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
2941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
3001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
3061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
3121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
3181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
3241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
3301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
3361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
3421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
3481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
3541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
3601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
3661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
3721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
3781 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
3841 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
3901 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
3961 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
4021 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
4081 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
4141 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
4201 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
4261 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
4321 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
4381 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
4441 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
4501 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
4561 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
4621 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttggtac
4681 cgagctcgga tccactagta acggccgcca gtgtgctgga attcctcgaa agacaactca
4741 gagttcacca tgggctccat cggtgcagca agcatggaat tttgttttga tgtattcaag
4801 gagctcaaag tccaccatgc caatgagaac atcttctact gccccattgc catcatgtca
4861 gctctagcca tggtatacct gggtgcaaaa gacagcacca ggacacaaat aaataaggtt
4921 gttcgctttg ataaacttcc aggattcgga gacagtattg aagctcagtg tggcacatct
4981 gtaaacgttc actcttcact tagagacatc ctcaaccaaa tcaccaaacc aaatgatgtt
5041 tattcgttca gccttgccag tagactttat gctgaagaga gatacccaat cctgccagaa
5101 tacttgcagt gtgtgaagga actgtataga ggaggcttgg aacctatcaa ctttcaaaca
5161 gctgcagatc aagccagaga gctcatcaat tcctgggtag aaagtcagac aaatggaatt
5221 atcagaaatg tccttcagcc aagctccgtg gattctcaaa ctgcaatggt tctggttaat
5281 gccattgtct tcaaaggact gtgggagaaa gcatttaagg atgaagacac acaagcaatg
5341 cctttcagag tgactgagca agaaagcaaa cctgtgcaga tgatgtacca gattggttta
5401 tttagagtgg catcaatggc ttctgagaaa atgaagatcc tggagcttcc atttgccagt
5461 gggacaatga gcatgttggt gctgttgcct gatgaagtct caggccttga gcagcttgag
5521 agtataatca actttgaaaa actgactgaa tggaccagtt ctaatgttat ggaagagagg
5581 aagatcaaag tgtacttacc tcgcatgaag atggaggaaa aatacaacct cacatctgtc
5641 ttaatggcta tgggcattac tgacgtgttt agctcttcag ccaatctgtc tggcatctcc
5701 tcagcagaga gcctgaagat atctcaagct gtccatgcag cacatgcaga aatcaatgaa
5761 gcaggcagag aggtggtagg gtcagcagag gctggagtgg atgctgcaag cgtctctgaa
5821 gaatttaggg ctgaccatcc attcctcttc tgtatcaagc acatcgcaac caacgccgtt
5881 ctcttctttg gcagatgtgt ttccccttaa aaagaagaaa gctgaaaaac tctgtccctt
5941 ccaacaagac ccagagcact gtagtatcag gggtaaaatg aaaagtatgt tatctgctgc
6001 atccagactt cataaaagct ggagcttaat ctagagggcc ctattctata gtgtcaccta
6061 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
6121 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
6181 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
6241 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
6301 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
6361 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
6421 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
6481 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
6541 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
6601 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
6661 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt t
//
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