| 质粒名称: | hTLR2 flag |
|---|---|
| 出品公司: | Sigma |
| 目录编号: | 13082 |
| 存储实验室: | Ruslan Medzhitov |
|
载体骨架基本信息 |
|
| 载体名称: | pFlag-CMV-1 |
| 载体出品公司: | Sigma |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 4989 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | TLR2 / toll-like receptor 2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2300 bp |
| 基因突变: | Amino acids 1-16 have been replaced with mammalian signal sequence (SALLILALVGAAVA) followed by Flag tag |
| GenBank编号 | NM_003264 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII(未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | CMV30 |
| 3’测序引物 | CMV24 |
|
文献引用方法 |
|
| 材料和方法部分: | hTLR2 flag was a gift from Ruslan Medzhitov (Addgene plasmid # 13082 ; http://n2t.net/addgene:13082 ; RRID:Addgene_13082) |
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 hTLR2 flag 7017 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS hTLR2 flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7017)
TITLE toll-like receptors
REFERENCE 2 (bases 1 to 7017)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7017
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 235..254
/label=T7
/note="T7 promoter, forward primer"
promoter 235..253
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(292..621)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 306..441
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(360..379)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 606..626
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
polyA_signal 650..1272
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind 1054..1073
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
CDS complement(3630..3653)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS complement(3654..3698)
/codon_start=1
/product="leader sequence from mouse preprotrypsin"
/label=preprotrypsin leader
/translation="MSALLILALVGAAVA"
promoter complement(3729..3932)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3758..3778)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3933..4312
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(4473..4490)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4473..4489)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4482..4504)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(4630..5085)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4767..4788)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4979..4998
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5112..5216
/gene="bla"
/label=AmpR promoter
CDS 5217..6077
/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(5435..5454)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6248..6836
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6737..6756
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6990..7007
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
1261 gggatgccac ccgggatcct ctagatccca actagacaaa gactggtctt aaatatggga
1321 acctaggact ttatcgcagc tctcagattt acccaaaatc cttcccgctg agcctcgtcc
1381 atgggccact ccaggtaggt cttggtgttc attatcttcc gcagcttgca gaagcgctgg
1441 ggaatggctt ttttctcaat gggctccaga agaatgagaa tggcagcatc attgttctca
1501 tcaaaaagac ggaaatggga gaagtccagt tcatacttgc accactcact cttcacaaag
1561 ttttcagaaa gcacaaagac agttttgtgg ctcttttcaa tggagtcaat gatattgtca
1621 atgatccact tgccaggaat gaagtcccgc ttatgaagac acaacttgaa ggggggattg
1681 aagttctcca gctcctggac cataaggttc tccacccagt aggcatcccg ctcactgtaa
1741 gaaacaaatg catcatagca gatgttcctg ctgggagctt tcctgggctt ccttttggcc
1801 tggagccagg cccacatcat tttcatatac cacaggccat ggaaacggtg gcacaggacc
1861 cccgtgagca ggatcagcag gaacagagca cagcacatgc cagacaccag tgctgtcctg
1921 tgacattccg acaccgagag gcggacatcc tgaacctgct ggccacgcac atgggatgga
1981 gagtcacaca ggtaatttgc tggccaatca atcaagactt tggccagtgc ttgctgctcc
2041 tgagtgaagg agaggaattc acaggagcaa atgaagttat tgccaccagc ttccaaagtc
2101 ttcagtgtgt gaaatgagtc aagttgctcc ttagaaaacg tagttattgc attcctactg
2161 attttcaata ctagtaacat gggtaagagg gaggcatctg gtagagtcat caacttattt
2221 ctggaaatat aaagttcttt gagttgcggc aaattcaaag aaaataaatt gagattgttg
2281 ttgctaacat ctaaaatttc cagtgtcttg ggaatgcagc ctgttacact gtgtattcgt
2341 gtgctggata agttcaaata tttcatcttt tctggccact gacaagtttc aggcatagaa
2401 tgaaaactat tcttactgat atcaatgtta gtcaagtttt tcagagtgag caaagtctct
2461 ccggtttttt ccaatgatgc caaatgattt tgccttaaaa ttaaagtttg tagagagggc
2521 caggcatcct cacaggctga atttttcaag tattcttcaa ccatcaaatt ttcactgaga
2581 tccaagtatt ctaatgattt taaatgttgt gaaagtaaac aaggaaccag aaaaacttta
2641 ctgttttcta ctgtgattct tttaactctt tctgtaagtg aatataaagt gctcagatca
2701 taaaataagt aaaaccttgg aatatgcagc ctccggattg ttaacgtttc cactttacct
2761 ggatctataa ctctgtcatt atcagatgct ctaaaattac caactccatt aagggtacag
2821 tcatcaaact ctaattctaa caatccagaa atctgattca aaagtttcat aacctgaaac
2881 aaactttcat cggtgatttt cacatttcta aatgtaaact ttttaatcaa tgaatttgtt
2941 tcaccagtgg atagttctga aaaatggaaa gtgtccaaat cagtatctcg cagttccaaa
3001 cattccacgg aacttgtaac atctacaaaa atctccagca gtaaaatatg ctgcttcata
3061 tgaaggatca gatgacttac attctgaatt gacttcaaac tttttggctc atagctctgt
3121 agatctgaag catcaatctc aagttcctca aggaaggtaa gtccagcaaa atcttttctt
3181 tgaatcttag tgaaggtgtc catatttccc actctcagga tttgcaattt tgtgagatga
3241 gaaaaaagag atgtttcccc tagggttttg taaggatttc ccagtaagtt taagaatgtt
3301 aaagaagaaa ggggcttgaa ccaggaagac gataaattag ataagtaatt ataggataag
3361 tctaaatgtt caagactgcc cagggaagaa aaagaatctt cctctattgt gttaattcca
3421 ttggatgtca gcaccagagc ctggaggttc acacacctct gtaggtcact gttgctaatg
3481 taggtgatcc tgttgttgga caggtcaagg ctttttacag cttctgtgag ccctgaggga
3541 atggagttta aagatcctga gctgcccttg cagataccat tgcggtcaca agacagagaa
3601 gcctgattgg aggattcttc cttggaaagc ttgtcgtcat cgtctttgta gtcagcaact
3661 gcagctccaa caagagctag gatcagaagt gcagacatgg tgaattaatt ctgacggttc
3721 actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
3781 aacggggcgg ggttattacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
3841 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
3901 cgcccattgg tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
3961 atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact gggcataatg ccaggcgggc
4021 catttaccgt cattgacgtc aatagggggc ggacttggca tatgatacac ttgatgtact
4081 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
4141 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
4201 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
4261 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacatggc
4321 ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc
4381 aatggccaat agccaatatt gatttatgct atataaccaa tgactaatat ggctaattgc
4441 caatattgat tcaatgtata gatcagcttg gcactggccg tcgttttaca acgtcgtgac
4501 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
4561 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
4621 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
4681 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
4741 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
4801 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
4861 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
4921 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
4981 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
5041 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
5101 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
5161 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
5221 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
5281 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
5341 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
5401 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
5461 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
5521 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
5581 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
5641 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
5701 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
5761 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
5821 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
5881 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
5941 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
6001 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
6061 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
6121 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
6181 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
6241 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
6301 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
6361 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
6421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
6481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
6541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
6601 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
6661 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
6721 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
6781 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
6841 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
6901 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
6961 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
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