| 质粒名称: | Stat3 Flag pRc/CMV |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 8707 |
| 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 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2600 bp |
| 基因突变: | *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(未破坏) |
| 3' 克隆位点: | ApaI (未破坏) |
| 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 Flag pRc/CMV was a gift from Jim Darnell (Addgene plasmid # 8707 ; http://n2t.net/addgene:8707 ; RRID:Addgene_8707) |
| 参考文献部分: | A STAT protein domain that determines DNA sequence recognition suggests a novel DNA-binding domain. Horvath CM, Wen Z, Darnell JE Jr. Genes Dev 1995 Apr 15;9(8):984-94. 10.1101/gad.9.8.984 PubMed 7774815 |
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 Flag pRc/CMV 7969 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Stat3 Flag pRc/CMV
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7969)
AUTHORS Horvath CM, Wen Z, Darnell JE Jr
TITLE A STAT protein domain that determines DNA sequence recognition
suggests a novel DNA-binding domain.
JOURNAL Genes Dev 1995 Apr 15;9(8):984-94.
PUBMED 7774815
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 103..120
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 237..258
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 273..303
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 311..327
/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 316..338
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 335..351
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 335..351
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 382..516
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(406..425)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 460..479
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(690..1484)
/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(801..820)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 1411..1430
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(1551..1880)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1565..1700
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1619..1638)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1865..1885
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(1908..2421)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2103..2124)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2315..2334
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 2467..2691
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 2680..2697
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 2717..2735
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2717..2734
/label=SP6
/note="SP6 promoter, forward primer"
CDS complement(2744..2767)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(5175..5191)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5175..5191)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(5281..5300)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5282..5300)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(5345..5548)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(5374..5394)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 5549..5928
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 6100..6119
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 6194..6298
/gene="bla"
/label=AmpR promoter
CDS 6299..7159
/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(6517..6536)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7330..7918
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7819..7838
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
61 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
121 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
241 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
301 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
361 gccaagctct cgaggtcgac ggatccagac atgataagat acattgatga gtttggacaa
421 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
481 ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat
541 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
601 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
661 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
721 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
781 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
841 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
901 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
961 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
1021 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
1081 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
1141 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
1201 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
1261 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
1321 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
1381 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
1441 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
1501 cctgtctctt gatcagatcc gaaaatggat atccaagctc ccgggagctt tttgcaaaag
1561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
1621 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
1681 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
1741 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
1801 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
1861 ccacacccta actgacacac attccacaga attcgagctc ggtacccacc ccggttgata
1921 atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaagcgtta
1981 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
2041 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
2101 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
2161 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
2221 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
2281 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
2341 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
2401 atgcgccgct acagggcgcg tggggatccc ccctcgagcc ccagctggtt ctttccgcct
2461 cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc
2521 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
2581 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
2641 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg
2701 atcagcgagc tctagcattt aggtgacact atagaatagg gcccttgtca tcgtcgtcct
2761 tgtagtctct agacatgggg gaggtagcac actccgaggt cagatccatg tcaaacgtga
2821 gcgactcaaa ctgccctcct gctgagggct cagcaccttc accgttattt ccaaactgca
2881 tcaatgaatc taaagtgcgg ggggacatcg gcaggtcaat ggtattgctg caggtcgttg
2941 gtgtcacaca gatgaacttg gtcttcaggt acggggcagc actacctggg tcggcttcgg
3001 ggtgctcctg gctctcgggc ctacagtact ttccaaatgc ctcctccttg ggaatgtcgg
3061 ggtagaggta gacaagtgga gacaccagga tgttggtcgc atccatgatc ttatagccca
3121 tgatgatttc agcaaatgac atgttgttca gctgctgctt ggtgtatggc tctacagact
3181 ggatctgggt cttgccactg atgtcctttt ccacccaagt gaaagtgacc cctccttctt
3241 tgctgctctc gctgaagcgc agtaggaagg tgcccggggg ctttgtgctt aggatggccc
3301 gctcccgctc cttgctgatg aaacccatga tgtacccttc attccaaagg gccaagatat
3361 actttttcac aaggtcgatg atattgtcta gccagaccca gaaggagaag cccttgccag
3421 ccatgttttc tttgcagaat ttagcccatg tgatctgaca ccctgagtag ttcacaccag
3481 gccctaggag cttctcagcc agcgttgtca gctgctcgat gctcagccct cgcttggtgg
3541 tggacgagaa ctgccagctg agcacctcgg ccacttggtc ccaggttcca attggcggct
3601 tagtgaagaa gttcacgttc ttggggttat tggtcagcat gttataccac aggattgatg
3661 cccaagcatt tggcatctga cagatgttgg agatcaccac aactggcaag gagtgggtct
3721 ctaggtcaat cttgaggcct tggtggtaca cctcagtctc gaaggtgatc aggtgcagct
3781 cctcagtcac gatcaaggag gcatcacaat tggcacggcc tccattccca catctctgct
3841 ccctaagggt caggtgcttg aactctgcag acaggctgcc gttgttagac tcctccatgt
3901 tcatcacttt tgtgttcgtg cccagaatgt taaatttccg agaccctctg agggcagcaa
3961 catccccaga gtctttatca atgcacactt taattttaag ctgataattc aactcaggaa
4021 atttgaccag caacctgact ttcgtggtaa actggacacc agtcttgatg actaagggcc
4081 ggtccgggtg catgggcatg cagggctgcc gctccaccac gaaggcactc ttcattaagt
4141 ttctgaacag ctccacgatc ctctcctcca gcatgggccg gtgctgcacg atagggtcgc
4201 ccttgtagga cactttctgc tgcagctcct ccagtttctt aatttgttgg cgggtctgaa
4261 gttgagattc tgctaatgaa gttatccagt tttccagacg gtccaggcag atgttgggag
4321 ggcctccgat gcacgcgatc tgctgccgcc tcttccagtc agccagctct tcatcagtca
4381 gtgtcttctg cacgtactcc attgctgaca agagccccgc cagctcactc acaatgcttc
4441 tccgcatctg gtccagggct gtgagcatct gttccagctg ctgcatcttc tgtctggtca
4501 cagactggtt gtttccattc agatcctgca tgtctccttg gctcttgagg gttttgtagt
4561 tgaaatcaaa gtcgtcctgg aggttctcca ccaccttcat tttctgttct agatcctgca
4621 ctcgcttccg gacatcctga agatgctgct ccaacatctg ctgcttctct gtcactacgg
4681 cggctgttgg gtggttggcc tggccccctt gctgggctgc cgtggctgcc gtctggagga
4741 ggcgagactc ttcccacagg catcgggcca cgatccgggc aatttccatt ggcttctcaa
4801 gatacctgct ctgcagaaac tgcttgattc ttcgaaggtt gtgctgatag aggacattgg
4861 actcttgcag gaatcggcta tattgctggt caatttcacc caagagatta tgaaacacca
4921 acgtggcatg tgactctttg ctggctgcat atgcccagtc ttgactctca atccaaggtg
4981 ccaggaactg ccgcagctcc atggggaagc tgtcgctgta cagctggtgc agctgcttca
5041 ggtagcgtgt gtccagctgc tgcagctggt tccactgagc catcctgctg cagtcagggg
5101 tctcgactgt ctccggggct gaggcccgac tggccggcct gcgctggtcg cggcggaatt
5161 cctgcagccc gggggatcca ctagttctag agcggccgcc agtgtgatgg atatctgcag
5221 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgggtct
5281 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
5341 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
5401 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
5461 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
5521 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
5581 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
5641 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
5701 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
5761 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
5821 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
5881 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
5941 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
6001 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
6061 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
6121 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
6181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
6241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
6301 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
6361 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
6421 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
6481 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
6541 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
6601 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
6661 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
6721 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
6781 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
6841 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
6901 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
6961 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
7021 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
7081 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
7141 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
7201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
7261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
7321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
7381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
7441 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
7501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
7561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
7621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
7681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
7741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
7801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
7861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
7921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggcctttt
//
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