| 质粒名称: | pcDNA3.1(-) HuTF-Fc-His tag |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 183253 |
| 存储实验室: | Alexander McLellan |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 6849bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Tissue factor |
| 物种来源: | H. sapiens (human) |
| 标签/融合蛋白: | 8xHis (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1(-) HuTF-Fc-His tag was a gift from Alexander McLellan (Addgene plasmid # 183253 ; http://n2t.net/addgene:183253 ; RRID:Addgene_183253) |
买家导航
pcDNA3.1(-) HuTF-Fc-His tag载体质粒基本信息
pcDNA3.1(-) HuTF-Fc-His tag质粒图谱载体图谱和 pcDNA3.1(-) HuTF-Fc-His tag载体序列质粒序列多克隆位点信息
 HuTF-Fc-His tag质粒图谱.png)
pcDNA3.1(-) HuTF-Fc-His tag质粒载体简介
pcDNA3.1(-) HuTF-Fc-His tag质粒序列载体序列
LOCUS pcDNA3.1(-) HuTF-Fc-His tag 6849 bp DNA circular SYN 15-APR-2025
DEFINITION Soluble expression of the extracellular domains of human tissue
factor (TF)-Fc.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6849)
TITLE McLellan Lab Plasmids 2022
REFERENCE 2 (bases 1 to 6849)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Sequence Label: pcDNA3.1(-) HuTF-Fc-His tag
FEATURES Location/Qualifiers
source 1..6849
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(26..43)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 32..256
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 302..730
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(389..408)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 599..620
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(739..759)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 744..1073
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 924..1059
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 986..1005
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1140..1934
/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(1194..1213)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1804..1823
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 2108..2229
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2145..2164)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2199..2218
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2278..2294)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2278..2294)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2291..2313)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2302..2318
/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(join(2326..2332,2333..2350,2351..2356))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2371..2392
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2509..2526)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2680..3268)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2760..2779)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(3439..4230,4231..4299))
/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 4062..4081
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4300..4404)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4479..4498)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4670..5049
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5050..5253
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5204..5224
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5298..5317
/label=T7
/note="T7 promoter, forward primer"
promoter 5298..5316
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 6809..6832
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
ORIGIN
1 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
61 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
121 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
181 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
241 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
301 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
481 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
601 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
721 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
901 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
961 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
1021 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
1081 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
1141 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
1201 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
1261 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
1321 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
1381 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
1441 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
1501 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
1561 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
1621 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
1681 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
1741 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
1801 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
1861 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
1921 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
1981 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
2041 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
2101 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
2161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
2221 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
2281 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
2341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
2401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
2461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
2521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
3481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
3541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
3601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
3661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
3721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
3781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
3841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
3901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
3961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
4021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
4081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
4141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
4201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
4261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
4381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
4441 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
4501 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
4561 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
4621 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
4681 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
4741 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
4801 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
4861 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
4921 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
4981 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
5041 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
5101 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
5161 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
5221 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
5281 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
5341 aacgggccct ctagactcga gcggccgcca ctgtgctgga tatctgcaga attcggcctc
5401 tgaggccacc atgcgaagga tgcaacttct tctgctcatc gcactttcct tggcgctggt
5461 gacaaatagt actacaaata ctgtggcagc atataattta acttggaaat caactaattt
5521 caagacaatt ttggagtggg aacccaaacc cgtcaatcaa gtctacactg ttcaaataag
5581 cactaagtca ggagattgga aaagcaaatg cttttacaca acagacacag agtgtgacct
5641 caccgacgag attgtgaagg atgtgaagca gacgtacttg gcacgggtct tctcctaccc
5701 ggcagggaat gtggagagca ccggttctgc tggggagcct ctgtatgaga actccccaga
5761 gttcacacct tacctggaga caaacctcgg acagccaaca attcagagtt ttgaacaggt
5821 gggaacaaaa gtgaatgtga ccgtagaaga tgaacggact ttagtcagaa ggaacaacac
5881 tttcctaagc ctccgggatg tttttggcaa ggacttaatt tatacacttt attattggaa
5941 atcttcaagt tcaggaaaga aaacagccaa aacaaacact aatgagtttt tgattgatgt
6001 ggataaagga gaaaactact gtttcagtgt tcaagcagtg attccctccc gaacagttaa
6061 ccggaagagt acagacagcc cggtagagtg tatgggccag gagaaagggg agttcagaga
6121 aggtaccgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
6181 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
6241 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
6301 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
6361 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
6421 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
6481 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
6541 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
6601 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
6661 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
6721 gcaggggaac gtcttctcat gctccgtgat gcacgaggct ctgcacaacc actacacgca
6781 gaagagcctc tccctgtctc cgggtaaaca tcaccaccat caccatcatc actgaggcct
6841 gtcaggcca
//
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