| 质粒名称: | Stat3-C Flag pRc/CMV |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 8722 |
| 存储实验室: | Jim Darnell |
|
载体骨架基本信息 |
|
| 载体名称: | pRc/CMV |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5500 bp |
| 完整质粒大小: | -- |
| 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Stat3-C,Stat3 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2600 bp |
| 基因突变: | A662C and N664C. Residues changed to Cys renders the molecule constitutively active. *See comments |
| GenBank编号 | U06922 |
| 启动子 | -- |
| 标签/融合蛋白: | Flag（插接件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NotI（未销毁） |
| 3' 克隆位点: | ApaI（未销毁） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: | Stat3-C Flag pRc/CMV was a gift from Jim Darnell (Addgene plasmid # 8722 ; http://n2t.net/addgene:8722 ; RRID:Addgene_8722) |
| 参考文献部分: | Stat3 as an oncogene. Bromberg JF, Wrzeszczynska MH, Devgan G, Zhao Y, Pestell RG, Albanese C, Darnell JE Jr. Cell 1999 Aug 6;98(3):295-303. 10.1016/S0092-8674(00)81959-5 PubMed 10458605 |
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 Stat3-C Flag pRc/CMV 7969 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Stat3-C Flag pRc/CMV
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7969)
AUTHORS Bromberg JF, Wrzeszczynska MH, Devgan G, Zhao Y, Pestell RG,
Albanese C, Darnell JE Jr
TITLE Stat3 as an oncogene.
JOURNAL Cell 1999 Aug 6;98(3):295-303.
PUBMED 10458605
REFERENCE 2 (bases 1 to 7969)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7969
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 296..315
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(534..638)
/gene="bla"
/label=AmpR promoter
primer_bind complement(713..732)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 904..1283
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1284..1487
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1438..1458
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1532..1551
/label=T7
/note="T7 promoter, forward primer"
promoter 1532..1550
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 1641..1657
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1641..1657
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 4065..4088
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(4097..4115)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4098..4115)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4135..4152)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4141..4365
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4411..4924
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4498..4517)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4708..4729
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4947..4967)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4952..5281
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5132..5267
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5194..5213
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5348..6142
/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(5402..5421)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6012..6031
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6316..6450
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6353..6372)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6407..6426
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6481..6497)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6481..6497)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6494..6516)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6505..6521
/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(6529..6559)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6574..6595
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6712..6729)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6883..7471)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6963..6982)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7642..533)
/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"
ORIGIN
1 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
61 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
661 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
721 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
781 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
841 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
901 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
961 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
1021 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
1081 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
1141 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
1201 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
1261 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
1321 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
1381 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
1441 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
1501 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
1561 cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctggaat tctgcagata
1621 tccatcacac tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattccgcc
1681 gcgaccagcg caggccggcc agtcgggcct cagccccgga gacagtcgag acccctgact
1741 gcagcaggat ggctcagtgg aaccagctgc agcagctgga cacacgctac ctgaagcagc
1801 tgcaccagct gtacagcgac agcttcccca tggagctgcg gcagttcctg gcaccttgga
1861 ttgagagtca agactgggca tatgcagcca gcaaagagtc acatgccacg ttggtgtttc
1921 ataatctctt gggtgaaatt gaccagcaat atagccgatt cctgcaagag tccaatgtcc
1981 tctatcagca caaccttcga agaatcaagc agtttctgca gagcaggtat cttgagaagc
2041 caatggaaat tgcccggatc gtggcccgat gcctgtggga agagtctcgc ctcctccaga
2101 cggcagccac ggcagcccag caagggggcc aggccaacca cccaacagcc gccgtagtga
2161 cagagaagca gcagatgttg gagcagcatc ttcaggatgt ccggaagcga gtgcaggatc
2221 tagaacagaa aatgaaggtg gtggagaacc tccaggacga ctttgatttc aactacaaaa
2281 ccctcaagag ccaaggagac atgcaggatc tgaatggaaa caaccagtct gtgaccagac
2341 agaagatgca gcagctggaa cagatgctca cagccctgga ccagatgcgg agaagcattg
2401 tgagtgagct ggcggggctc ttgtcagcaa tggagtacgt gcagaagaca ctgactgatg
2461 aagagctggc tgactggaag aggcggcagc agatcgcgtg catcggaggc cctcccaaca
2521 tctgcctgga ccgtctggaa aactggataa cttcattagc agaatctcaa cttcagaccc
2581 gccaacaaat taagaaactg gaggagctgc agcagaaagt gtcctacaag ggcgacccta
2641 tcgtgcagca ccggcccatg ctggaggaga ggatcgtgga gctgttcaga aacttaatga
2701 agagtgcctt cgtggtggag cggcagccct gcatgcccat gcacccggac cggcccttag
2761 tcatcaagac tggtgtccag tttaccacga aagtcaggtt gctggtcaaa tttcctgagt
2821 tgaattatca gcttaaaatt aaagtgtgca ttgataaaga ctctggggat gttgctgccc
2881 tcagagggtc tcggaaattt aacattctgg gcacgaacac aaaagtgatg aacatggagg
2941 agtctaacaa cggcagcctg tctgcagagt tcaagcacct gacccttagg gagcagagat
3001 gtgggaatgg aggccgtgcc aattgtgatg cctccttgat cgtgactgag gagctgcacc
3061 tgatcacctt cgagactgag gtgtaccacc aaggcctcaa gattgaccta gagacccact
3121 ccttgccagt tgtggtgatc tccaacatct gtcagatgcc aaatgcttgg gcatcaatcc
3181 tgtggtataa catgctgacc aataacccca agaacgtgaa cttcttcact aagccgccaa
3241 ttggaacctg ggaccaagtg gccgaggtgc tcagctggca gttctcgtcc accaccaagc
3301 gagggctgag catcgagcag ctgacaacgc tggctgagaa gctcctaggg cctggtgtga
3361 actactcagg gtgtcagatc acatgggcta aattctgcaa agaaaacatg gctggcaagg
3421 gcttctcctt ctgggtctgg ctagacaata tcatcgacct tgtgaaaaag tatatcttgg
3481 ccctttggaa tgaagggtac atcatgggtt tcatcagcaa ggagcgggag cgggccatcc
3541 taagcacaaa gcccccgggc accttcctac tgcgcttcag cgagagcagc aaagaaggag
3601 gggtcacttt cacttgggtg gaaaaggaca tcagtggcaa gacccagatc cagtctgtag
3661 agccatacac caagcagcag ctgaacaaca tgtcatttgc tgaaatcatc atgggctata
3721 agatcatgga ttgtacctgc atcctggtgt ctccacttgt ctacctctac cccgacattc
3781 ccaaggagga ggcatttgga aagtactgta ggcccgagag ccaggagcac cccgaagccg
3841 acccaggtag tgctgccccg tacctgaaga ccaagttcat ctgtgtgaca ccaacgacct
3901 gcagcaatac cattgacctg ccgatgtccc cccgcacttt agattcattg atgcagtttg
3961 gaaataacgg tgaaggtgct gagccctcag caggagggca gtttgagtcg ctcacgtttg
4021 acatggatct gacctcggag tgtgctacct cccccatgtc tagagactac aaggacgacg
4081 atgacaaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
4141 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
4201 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
4261 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
4321 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
4381 gaaccagctg gggctcgagg ggggatcccc acgcgccctg tagcggcgca ttaagcgcgg
4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
4741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
4861 caatttaaat atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg
4921 gggtgggtac cgagctcgaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
4981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
5041 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
5101 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
5161 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
5221 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
5281 aaagctcccg ggagcttgga tatccatttt cggatctgat caagagacag gatgaggatc
5341 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
5401 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
5461 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
5521 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
5581 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
5641 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
5701 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
5761 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
5821 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
5881 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
5941 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
6001 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
6061 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
6121 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
6181 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
6241 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
6301 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
6421 ctcatcaatg tatcttatca tgtctggatc cgtcgacctc gagagcttgg cgtaatcatg
6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
6541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
7321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtg
//
