| 质粒名称: | pUB-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 11155 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 3084 bp |
| 完整质粒大小: | 4295 bp |
| 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: | Ubiquitin C promoter |
| 物种来源: | 人源 |
| 插入大小: | 1211 bp |
| 基因又名: | UBC；UBC promoter |
| 标签/融合蛋白: | EGFP (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SpeI(未破坏) |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | na |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pUB-GFP was a gift from Connie Cepko (爱迪基因 plasmid # 11155) |
| 参考文献部分: | Electroporation and RNA interference in the rodent retina in vivo and in vitro. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. 10.1073/pnas.2235688100 PubMed 14603031 |
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载体基本信息
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载体简介
载体序列
LOCUS pUB-GFP 5125 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression vector for expression of GFP (Ubiquitin C
promoter).
ACCESSION .
VERSION .
KEYWORDS pUB-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5125)
AUTHORS Matsuda T, Cepko CL
TITLE Electroporation and RNA interference in the rodent retina in vivo
and in vitro.
JOURNAL Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
PUBMED 14603031
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pUB-GFP|
FEATURES Location/Qualifiers
source 1..5125
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 116..135
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 375..396
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 438..447
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(524..1735)
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind complement(668..687)
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
promoter 1808..1912
/gene="bla"
/label=AmpR promoter
CDS 1913..2773
/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(2131..2150)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2944..3532
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3433..3452
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3686..3703
/label=L4440
/note="L4440 vector, forward primer"
polyA_signal 3771..3905
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3795..3814)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3849..3868
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter complement(3911..4106)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3925..4060
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3979..3998)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 4165..4186
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4201..4231
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4239..4255
/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 4244..4266
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4263..4279
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4263..4279
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 4622..4641
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
polyA_site 4640..4695
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind 4741..4760
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
CDS complement(4847..441)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4892..4913)
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
61 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
121 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
181 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
241 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
301 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
361 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
421 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
481 ctgcagaatt cgaagcttga gctcgacctg cagcccaagc ttcgtctaac aaaaaagcca
541 aaaacggcca gaatttagcg gacaatttac tagtctaaca ctgaaaatta catattgacc
601 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg
661 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc
721 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca
781 ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac
841 ctaccggcag gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac
901 gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc
961 cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt
1021 ccccagatgg tgccccagcc catctcaccc gaataagagc tttcccgcat tagcgaaggc
1081 ctcaagacct tgggttcttg ccgcccacca tgccccccac cttgtttcaa cgacctcaca
1141 gcccgcctca caagcgtctt ccattcaaga ctcgggaaca gccgccattt tgctgcgctc
1201 cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc agactacagc ccttggcggt
1261 ctctccacac gcttccgtcc caccgagcgg cccggcggcc acgaaagccc cggccagccc
1321 agcagcccgc tactcaccaa gtgacgatca cagcgatcca caaacaagaa ccgcgaccca
1381 aatcccggct gcgacggaac tagctgtgcc acacccggcg cgtccttata taatcatcgg
1441 cgttcaccgc cccacggaga tccctccgca gaatcgccga gaagggacta cttttcctcg
1501 cctgttccgc tctctggaaa gaaaaccagt gccctagagt cacccaagtc ccgtcctaaa
1561 atgtccttct gctgatactg gggttctaag gccgagtctt atgagcagcg ggccgctgtc
1621 ctgagcgtcc gggcggaagg atcaggacgc tcgctgcgcc cttcgtctga cgtggcagcg
1681 ctcgccgtga ggaggggggc gcccgcggga ggcgccaaaa cccggcgcgg aggccagatc
1741 tggagccgac acgggttact agtcaataat caatgtcgac ccaggtggca cttttcgggg
1801 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
1861 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
1921 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
1981 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
2041 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
2101 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
2161 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
2221 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
2281 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
2341 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
2401 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
2461 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
2521 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
2581 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
2641 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
2701 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
2761 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
2821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
2881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
2941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
3001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
3061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
3121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
3181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
3241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
3301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
3361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
3421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
3481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
3541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
3601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
3661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
3721 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg gatccagaca
3781 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
3841 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
3901 aagttagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
3961 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
4021 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
4081 ctatggttgc tgactaattg agatgcggat ccgctggcac gacaggtttc ccgactggaa
4141 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
4201 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
4261 cacaggaaac agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc
4321 ataagagaag agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta
4381 gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata
4441 taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg acagcagggg
4501 gctgtttcat atactgatga cctctttata gccaaccttt gttcatggca gccagcatat
4561 gggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc
4621 tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa
4681 taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat
4741 ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcattg gccacaccag
4801 ccaccacctt ctgataggca gcctgcacct gaggagtgcg gccgctttac ttgtacagct
4861 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
4921 gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca
4981 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
5041 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
5101 cggccatgat atagacgttg tggct
//
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