| 质粒名称: | pcDNA3.1-TYMS |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 100545 |
| 存储实验室: | Yuh-Shan Jou |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA3.1+ |
| 载体出品公司: | invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5428 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: | thymidylate synthetase,TS, TMS,HST422, TYMS |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 942 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRV(未破坏) |
| 3' 克隆位点: | XhoI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1-TYMS was a gift from Yuh-Shan Jou (Addgene plasmid # 100545 ; http://n2t.net/addgene:100545 ; RRID:Addgene_100545) |
| 参考文献部分: | Pyrimidine metabolic rate limiting enzymes in poorly-differentiated hepatocellular carcinoma are signature genes of cancer stemness and associated with poor prognosis. Yeh HW, Lee SS, Chang CY, Hu CM, Jou YS. Oncotarget. 2017 Sep 8;8(44):77734-77751. doi: 10.18632/oncotarget.20774. eCollection 2017 Sep 29. 10.18632/oncotarget.20774 PubMed 29100421 |
买家导航
pcDNA3.1-TYMS载体 pcDNA3.1TYMS质粒基本信息
pcDNA3.1-TYMS质粒图谱 pcDNA3.1TYMS载体图谱和pcDNA3.1-TYMS载体序列 pcDNA3.1TYMS质粒序列多克隆位点信息
pcDNA3.1-TYMS质粒 pcDNA3.1TYMS载体简介
pcDNA3.1-TYMS质粒序列 pcDNA3.1TYMS载体序列
LOCUS pcDNA3.1-TYMS 6356 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION mammalian expression of thymidylate synthetase .
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1-TYMS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6356)
AUTHORS Yeh HW, Lee SS, Chang CY, Hu CM, Jou YS
TITLE Pyrimidine metabolic rate limiting enzymes in poorly-differentiated
hepatocellular carcinoma are signature genes of cancer stemness and
associated with poor prognosis.
JOURNAL Oncotarget. 2017 Sep 8;8(44):77734-77751. doi:
10.18632/oncotarget.20774. eCollection 2017 Sep 29.
PUBMED 29100421
REFERENCE 2 (bases 1 to 6356)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6356
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1713..1730)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1719..1943
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1989..2417
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2076..2095)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2286..2307
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2426..2446)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2431..2760
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2611..2746
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2673..2692
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2827..3621
/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(2881..2900)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3491..3510
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3795..3916
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3832..3851)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3886..3905
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3965..3981)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3965..3981)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3978..4000)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3989..4005
/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(4013..4043)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4058..4079
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4196..4213)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4367..4955)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4447..4466)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5126..5986)
/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 5749..5768
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5987..6091)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6166..6185)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
721 ttctgcagat atcatgcctg tggccggctc ggagctgccg cgccggccct tgccccccgc
781 cgcacaggag cgggacgccg agccgcgtcc gccgcacggg gagctgcagt acctggggca
841 gatccaacac atcctccgct gcggcgtcag gaaggacgac cgcacgggca ccggcaccct
901 gtcggtattc ggcatgcagg cgcgctacag cctgagagat gaattccctc tgctgacaac
961 caaacgtgtg ttctggaagg gtgttttgga ggagttgctg tggtttatca agggatccac
1021 aaatgctaaa gagctgtctt ccaagggagt gaaaatctgg gatgccaatg gatcccgaga
1081 ctttttggac agcctgggat tctccaccag agaagaaggg gacttgggcc cagtttatgg
1141 cttccagtgg aggcattttg gggcagaata cagagatatg gaatcagatt attcaggaca
1201 gggagttgac caactgcaaa gagtgattga caccatcaaa accaaccctg acgacagaag
1261 aatcatcatg tgcgcttgga atccaagaga tcttcctctg atggcgctgc ctccatgcca
1321 tgccctctgc cagttctatg tggtgaacag tgagctgtcc tgccagctgt accagagatc
1381 gggagacatg ggcctcggtg tgcctttcaa catcgccagc tacgccctgc tcacgtacat
1441 gattgcgcac atcacgggcc tgaagccagg tgactttata cacactttgg gagatgcaca
1501 tatttacctg aatcacatcg agccactgaa aattcagctt cagcgagaac ccagaccttt
1561 cccaaagctc aggattcttc gaaaagttga gaaaattgat gacttcaaag ctgaagactt
1621 tcagattgaa gggtacaatc cgcatccaac tattaaaatg gaaatggctg tttagctcga
1681 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
1741 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
1801 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
1861 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
1921 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
1981 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
2041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
2101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
2161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
2221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
2281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
2341 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
2401 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
2461 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
2521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2581 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
2641 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
2701 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
2761 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
2821 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
2881 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
2941 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
3001 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
3061 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
3121 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
3181 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
3241 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
3301 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
3361 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
3421 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
3481 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
3541 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
3601 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
3661 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
3721 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
3781 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
3841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
3901 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
3961 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
4021 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
4081 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
4141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
4201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
4261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
4321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
4381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
4441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
4501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
4561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
4621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
4681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
4741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
4801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
4861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
4921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
4981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
5041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
5101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
5161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
5221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
5281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
5341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
5401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
5461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
5521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
5581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
5641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
5701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
5761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
5821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
5881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
5941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
6001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
6061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
6121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
6181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
6241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
6301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
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