| 质粒名称: | pCralbp-DsRed |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 11158 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3086 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: | Cralbp promoter |
| 物种来源: | -- |
| 插入大小: | 4002 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: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pCralbp-DsRed was a gift from Connie Cepko (Addgene plasmid # 11158 ; http://n2t.net/addgene:11158 ; RRID:Addgene_11158) |
| 参考文献部分: | Electroporation and RNA interference in the rodent retina in vivo and in vitro. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. 10.1073/pnas.2235688100 PubMed 14603031 |
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载体基本信息
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载体简介
载体序列
LOCUS pCralbp-DsRed 7807 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCralbp-DsRed
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7807)
AUTHORS Matsuda T, Cepko CL
TITLE Electroporation and RNA interference in the rodent retina in vivo
and in vitro.
JOURNAL Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
PUBMED 14603031
REFERENCE 2 (bases 1 to 7807)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7807
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 931..939
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
regulatory 4063..4072
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4069..4746
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(4249..4269)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 4661..4684
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(4831..4850)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4896..4951
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4950..4969)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5310..5326)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5310..5326)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5323..5345)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5334..5350
/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(5358..5388)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5403..5424
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5483..5678
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5529..5664
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5591..5610
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5684..5818
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5721..5740)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5775..5794
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5886..5903)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6057..6645)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6137..6156)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6816..7676)
/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 7439..7458
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7677..7781)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agtgatccct ttgctagcac tgagaccctt taatacgcct
61 cctcatgttg tggggacccc ttaaccataa aattattttt gttgctccat cataactata
121 attttgtcac ctttatgaat cataatgtaa atatctgata ttcaggatat ctgatatgca
181 actcttgtga aagggtccat caaccacccc atcccccacc ctcagtgatc atgacccaca
241 ggttgggtct aggcagccct ctcctgtaag actctggcca gccttagctt acaggggatt
301 ccctgagtga acagcacaga gccgaaaatc gatgcaaaaa gcaagaggaa tttattgctc
361 caacatgttg gggtcgccct gcacatgaag agagagaccg accacaggca gcccttcctt
421 tacagcttct ccaggcagca gccattaggc acaatatgat tggcagaaca gtatgacttt
481 taaactgatt ggtctatagg gaatgaggta ggtggccaac agtatctctg agcctcccct
541 acccgcccgc aggtggccag tggttggtcc ctcctcccgg tctgaggaat gtaattagcc
601 ccttccttcc ggaggggatg ggtgcgttct ttaagacctt tctcttccag gaggggaggg
661 gtctggtgga attttctaaa gtacctgagc tgacctcttc actccaacta gaccatgtca
721 tggtagtcct cttggtacac agggatgctt ggtgatgaat cacagagcct cttagcagct
781 gggactctta cctcccatga gcatcacagc cagggaggtt ctttgggcat aacagcatgc
841 cacagcctag gtgaattaac caatagaaaa tcattctctc ggaatctcgg tagttggagg
901 tctaagatca aggtattatc ctttggtgtc acctccttat cttatagata aaccctgaac
961 tgtggctgga tttcagagaa cgacctgtgc acatgagctc aaccctaccc tgagcacatg
1021 gaggagagag aaccaggacc aaaggtgggc tgcttgggtc cagactgaag atgaatttaa
1081 ttacagacaa ctatctgtgg tggaagtagg gatctcccgg acattggagg cccccacaga
1141 gatctaaagg atgtgtatcc ctcatacccc ggtgataaac caaggcagag tctgggtgct
1201 cctttgactg ttggatgcat tcccttagcc tgggtgctca ctggctctcc attcactggg
1261 tacaactcag aactccccag aatacttccc agggattggg cagctaagcc cctgaagtgg
1321 gtactttgca tctcaggact ttccccacag cttcagctgg ctctctagtc atcccagttc
1381 tcagagggga ctgagtcatg tcgtggtgac ataaccacac tgaaaaaagt gcagtagcaa
1441 ggacagaaag tcggacaagg gagatctgct gaggccccac acccatcccc tgcttccttg
1501 cctttgcgag gtagagatcg gcagagatcg gagggtcagg gtacctagca agtcctccgc
1561 tgctcagcct cgcccgctcc atttccctct gccacatggg acagtggacc ataggcctgg
1621 gttgcagcaa tgatcacatt tataatttct cttcacttct ctatatgtcc tgccctttgt
1681 gaagaaggag tagtctccca ggcccccctg cctcccagtt tgagaagcga gatggcaggg
1741 aaatgagacg gggctggccc agatccagtg aagtggcttc cacatggctg aacagagagc
1801 tccaagcagg agctcacaaa gcagggcttt gatggtagcc ccgcaggccg ggcctctccc
1861 ctctccctca gggtgcttca cttgcttcat tctgggcctc ctgtgagcct gactgaatgg
1921 gaaacggtgg gcctgagaag ttccttgtga cacactaatc ccagcttgct gacctgacca
1981 cacccccaaa gaaagcagcc tcccgagctc cagaacaatt caggcatcca gtagccccca
2041 agaaagagct gtcaggtaag tcagtaacaa ggcctggctg ggccagtgag tataagaaag
2101 agtgtgtgca tgggagcggg gtaaatgatc agggatgggt ggcgaacacc acgtctcaga
2161 ttgcttggac cggcctgtaa aggcgtacgc catttccgca gggtgagaca gaccgatcca
2221 cgggcagacg caacacatac gaatgtggat tacctaactc aattagaatg gcaggtgatc
2281 aaatactctt tatgccctag tattggtaat taattgtact ttatttgcta attctaaatg
2341 ccggagaaca tcgatgtgga gataggggta gcatgttttt attacataca tgcctacagc
2401 taatgggtaa gaacattttt aaaataagag ataagtgctt ctccgtgctg acgttaatct
2461 tgcacttcta agatctggga cttgggctgc tgatctgcac atttggctag ggataatgca
2521 ctaatacctc tcgagtcttg ctttgctttg tttgcttgag gcagggtttc tctacgtagc
2581 cctagctatc ctggaactca ctatgaagaa cagactggct tcaaaatcag agattcccct
2641 agacactggg attaaagatg cgtaccacta tacccagctt ttgcttttcg tttttttgag
2701 atagagtctc atgtagcaca ggctggtttt gaattaacta tgtagccaag gataacctta
2761 actaaggatt ctcttgcctc tatctttttt tttctttcct ttttaaaaat tactagatat
2821 tttctttgtt tacatttcaa atgttatcgc ctttcctagt ttcctctcca aaagtcccct
2881 ataccctcac ccccgccctg ctccccaacc cacccatttc tgtttcctgg ccctggcatt
2941 cccctgtact ggggcatata atcttcgcaa gactaagggc ctctcctccc attgatggcc
3001 aactaggcca tcttctgcta cataagcagc tagagacaca agctctgggg ggtactggtt
3061 agttcatatt gttgttcctc ctatagggct gcagacccct tcagctcctt gggtactttc
3121 tctagctcct tcgttgggga ccctgtgttc catcctatag atgactgtga gcgtccactt
3181 ctgtatttgc caggcactgc ttttgcctct atcttgcaag ttctgggaat acagatgtgt
3241 gccacaacag tcaactcttg gtctttacac ccacatatct ggatgtctgt ctactatcca
3301 aagagatggc ttgtacttga ttcttgtggg tagatcccag cccccaagaa ttccctggtc
3361 ctttgggggc tgtggtaggg aagggatagg ccctcagata ttctctgaga tcctcaagtt
3421 tcagtgtaca aattgggtta atctcagtgc tagcaaaggg agaaaagact gttgtcgatc
3481 aggagtcctc tctcggttat gtcacagaac cctgtggtcc tcacagcggt cacttttggt
3541 agatattatt atccttcgtt ggctctcaat ctcagaagga ttaaatacct tgcctatggt
3601 cacacaggtg gtaaaagtca gattcaaatt cagatctctc taacctataa ggtttctgtc
3661 accagctccc ccagggactt tgcaaagaga tgacttggct tgaggcaaag agacttagtg
3721 tcaggtggat acaggttcaa gtcctagctt aaataccaac tatgcgtaag tgaccgagct
3781 gaccaatgga acctagccac aggcagaaac agaatgtaaa gatgttcaca gtcactgggg
3841 acatttaggt cttggaggag aaaatctgga aatgtgggtc atgatggata tctgttctgg
3901 tggcccagaa atctatgctg gtgtatatta tctaattaca tattaatcct tctatcccaa
3961 caagggacgg tcaagactcg gatgaaatca cacaacttgc aagtgtgaga gacagcattg
4021 caatccagga tcgaattccc gcgggcccgg gatccaccgg tcgccaccat ggcctcctcc
4081 gagaacgtca tcaccgagtt catgcgcttc aaggtgcgca tggagggcac cgtgaacggc
4141 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcca caacaccgtg
4201 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
4261 ttccagtacg gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag
4321 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
4381 gcgaccgtga cccaggactc ctccctgcag gacggctgct tcatctacaa ggtgaagttc
4441 atcggcgtga acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag
4501 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagac ccacaaggcc
4561 ctgaagctga aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag
4621 aagcccgtgc agctgcccgg ctactactac gtggacgcca agctggacat cacctcccac
4681 aacgaggact acaccatcgt ggagcagtac gagcgcaccg agggccgcca ccacctgttc
4741 ctgtagcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
4801 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
4861 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
4921 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
4981 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct
5041 gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
5101 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
5161 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
5221 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg
5281 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
5341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
5401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
5461 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
5521 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
5581 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
5641 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
5701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
5761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
5821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
5881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
5941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
6001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
6061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
6121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
6181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
6241 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
6301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
6361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
6421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
6481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
6541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
6601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
6661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
6721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
6781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
6841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
6901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
6961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
7021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
7081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
7141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
7201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
7261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
7321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
7381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
7441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
7501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
7561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
7621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
7681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
7741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
7801 ccacctg
//
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