| 质粒名称: | pmRFP_LAP2beta_IRES_puro2b |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 21047 |
| 存储实验室: | Daniel Gerlich |
|
载体骨架基本信息 |
|
| 载体名称: | IRES_puro2b |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，细菌表达 |
| 筛选标记: | 嘌呤霉素 |
| 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: | LAP2_beta,Tmpo |
| 物种来源: | R. norvegicus (rat) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | mRFP |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pmRFP_LAP2beta_IRES_puro2b was a gift from Daniel Gerlich (Addgene plasmid # 21047 ; http://n2t.net/addgene:21047 ; RRID:Addgene_21047) |
| 参考文献部分: | Aurora B-mediated abscission checkpoint protects against tetraploidization. Steigemann P, Wurzenberger C, Schmitz MH, Held M, Guizetti J, Maar S, Gerlich DW. Cell. 2009 Feb 6. 136(3):473-84. 10.1016/j.cell.2008.12.020 PubMed 19203582 |
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 pmRFP_LAP2beta_IRES_puro2b 6414 bp ds-DNA circular SYN 21-FEB-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pmRFP_LAP2beta_IRES_puro2b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6414)
AUTHORS Steigemann P, Wurzenberger C, Schmitz MH, Held M, Guizetti J, Maar
S, Gerlich DW
TITLE Aurora B-mediated abscission checkpoint protects against
tetraploidization.
JOURNAL Cell. 2009 Feb 6. 136(3):473-84.
PUBMED 19203582
REFERENCE 2 (bases 1 to 6414)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6414
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 859..868
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 865..1539
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(995..1013)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1263..1282
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 1457..1480
/label=DsRed1-C
/note="DsRed1, forward primer"
intron 2277..2506
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 2489..2508
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 2564..3135
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2729..2746)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2956..2975
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3168..3767
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3168..3187)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3664..3684
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(3903..3920)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3909..4133
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 4180..4199
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4259..4275)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4259..4275)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4272..4294)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4283..4299
/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(4307..4337)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4352..4373
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4490..4507)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4661..5249)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4741..4760)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5420..6280)
/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 6043..6062
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6281..6385)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
841 ccgctagcgc taccggtcgc caccatggcc tcctccgagg acgtcatcaa ggagttcatg
901 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
961 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
1021 cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc caaggcctac
1081 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
1141 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
1201 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
1261 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg gatgtacccc
1321 gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga cggcggccac
1381 tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct gcccggcgcc
1441 tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
1501 cagtacgagc gcgccgaggg ccgccactcc accggcgccc tgtacaagta ctcagatctc
1561 gagctcaagc ttgcattaac tagggagtcc acgagagggt cgagaagaac tccaaggaga
1621 agggtggaac cctcacagca ttttcgtgta gatggtgcag taatttcaga gagtactccc
1681 atagctgaga ctataaaggc ttccagcaac gactccttag tggccaatag gttgactgga
1741 aatttcaagc atgcatcttc tattctgcca atcactgaat tctcagacat aaccagaaga
1801 acaccaaaga aaccattgac aagagctgaa gtgggagaaa aaacagagga aagaagagta
1861 gaaagggata ttctgaagga aatgttcccg tatgaagcct ctactccaac cggaattagt
1921 gctagttgcc gcagaccaat caaaggtgct gccggccggc cgctcgagct cagtgacttc
1981 aggatggaag aatcgttctc atctaagtac gtcccgaagt acgttccctt ggcagacgtc
2041 aagtcggaaa agacaaagaa gggacgctcc gttcccatgt ggataaaaat gctgctgttt
2101 gctctcgtgg cgggtttttt gtttttggtc tatcaagcta tggaaaccaa ccaaggaaac
2161 cccttcacta attttcttca agatactaaa atatccaact gaggatccgc ggccgcatag
2221 ataactgatc cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga
2281 gtactccctc tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga
2341 ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg
2401 gtcagaaaag acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc
2461 atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag
2521 gtccaactgc aggtcgagca tgcatctagg gcggccaatt ccgcccctct cccccccccc
2581 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
2641 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
2701 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
2761 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
2821 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
2881 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
2941 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
3001 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
3061 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
3121 aaaacacgat gataagcttg ccacaaccca caaggagacg accttccatg accgagtaca
3181 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
3241 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
3301 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
3361 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
3421 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
3481 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
3541 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
3601 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
3661 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
3721 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
3781 acccgcagcg cccgaccgaa aggagcgcac gaccccatgg ctccgaccga agccacccgg
3841 ggcggccccg ccgaccccgc acccgccccc gaggcccacc gactctagag ctcgctgatc
3901 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
3961 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
4021 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
4081 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
4141 ggcggaaaga accagctggg gctcgagtgc attctagttg tggtttgtcc aaactcatca
4201 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
4261 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
4321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
4501 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
4561 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
4621 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
4681 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
4741 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4801 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
4861 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
4921 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
4981 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
5041 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
5101 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
5161 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
5221 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
5281 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
5341 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
5401 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
5461 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
5521 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
5581 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
5641 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
5701 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
5761 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5821 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5881 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5941 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
6001 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
6061 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
6121 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
6181 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
6241 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
6301 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
6361 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacg
//
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