| 质粒名称: | pCMV4-HA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 27553 |
| 存储实验室: | Shao-Cong Sun |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV4 |
| 载体出品公司: | Dr. David Russell, UTSW |
| 空载体大小: | 4874 bp |
| 完整质粒大小: | 4970 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: | 无 |
| 标签/融合蛋白: | 3x HA |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NA (unknown if destroyed) |
| 3’克隆位点: | NA (unknown if destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | hGH-pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV4-HA was a gift from Shao-Cong Sun (Addgene plasmid # 27553 ; http://n2t.net/addgene:27553 ; RRID:Addgene_27553) |
| 参考文献部分: | Negative regulation of the nuclear factor kappa B-inducing kinase by a cis-acting domain. Xiao G, Sun SC. J Biol Chem. 2000 Jul 14. 275(28):21081-5. 10.1074/jbc.M002552200 PubMed 10887201 |
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 pCMV4-HA 4970 bp ds-DNA circular SYN 01-FEB-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCMV4-HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4970)
AUTHORS Xiao G, Sun SC
TITLE Negative regulation of the nuclear factor kappa B-inducing kinase by
a cis-acting domain.
JOURNAL J Biol Chem. 2000 Jul 14. 275(28):21081-5.
PUBMED 10887201
REFERENCE 2 (bases 1 to 4970)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 1, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4970
/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"
CDS 682..771
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
polyA_signal 821..1443
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(1020..1039)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(1467..1487)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1472..1801
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1652..1787
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1714..1733
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 1840..1936
/label=SV40 intron
/note="modified SV40 intron with splice donor and acceptor
sites"
primer_bind complement(2012..2031)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2013..2031)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2045..2061)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2045..2061)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2058..2080)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2069..2085
/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(2093..2123)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2138..2159
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2276..2293)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2447..3035)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2527..2546)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3206..4066)
/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 3829..3848
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4067..4171)
/gene="bla"
/label=AmpR promoter
rep_origin 4198..4653
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4285..4304)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4495..4516
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4779..4801
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4793..4810
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4794..4810
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agaattgttt ttatttttaa ttttctttca aatacttcca tcgaattcag atctggtacc
661 acgcgtacca tggatatctt ttacccatac gatgttcctg actatgcggg ctatccctat
721 gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc tgctcagatc
781 gataagcttg catgcctgca ggtcgactct agaggatccc gggtggcatc cctgtgaccc
841 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
901 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
961 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
1021 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
1081 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
1141 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
1201 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
1261 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
1321 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
1381 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
1441 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
1501 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
1561 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
1621 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
1681 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
1741 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
1801 aaagctcctc gaggaactga aaaaccagaa agttaactgg taagtttagt ctttttgtct
1861 tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg
1921 ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc tgctgcaggt
1981 cgactctagg gccgcgtacc gagctcgaat tccctatagt gagtcgtatt aaattcgtaa
2041 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
2101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
2161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
2221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
2281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
2341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
2401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
2461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
2521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
2581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
2641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
2701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
2761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
2821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
2881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
2941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
3001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
3061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
3121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
3181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
3241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
3301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
3361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
3421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
3481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
3541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
3601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
3661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
3721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
3781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
3841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
3901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
3961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
4021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
4081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
4141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
4201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
4261 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
4321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
4381 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
4441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
4501 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
4561 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
4621 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
4681 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
4741 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
4801 gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt
4861 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
4921 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
//
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