| 质粒名称: | |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 24218 |
| 存储实验室: | Mariano Garcia-Blanco |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA6/V5/His |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5002 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: | 杀螨素 |
| 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: | RFP with modified adeno intron,RFP splicing reporter |
| 物种来源: | synthetic and adenovirus |
| 插入大小: | 815 bp |
| 基因突变: | dsRED interrupted by a modified adenovirus intron |
| 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: | HindIII (未破坏) |
| 3' 克隆位点: | AgeI (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRint was a gift from Mariano Garcia-Blanco (Addgene plasmid # 24218 ; http://n2t.net/addgene:24218 ; RRID:Addgene_24218) |
| 参考文献部分: | A protocol for imaging alternative splicing regulation in vivo using fluorescence reporters in transgenic mice. Bonano VI, Oltean S, Garcia-Blanco MA. Nat Protoc. 2007 . 2(9):2166-81. 10.1038/nprot.2007.292 PubMed 17853873 |
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 pRint 5816 bp ds-DNA circular SYN 09-APR-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRint
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5816)
AUTHORS Bonano VI, Oltean S, Garcia-Blanco MA
TITLE A protocol for imaging alternative splicing regulation in vivo using
fluorescence reporters in transgenic mice.
JOURNAL Nat Protoc. 2007 . 2(9):2166-81.
PUBMED 17853873
REFERENCE 2 (bases 1 to 5816)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 9, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5816
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 918..927
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1053..1071)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind complement(1103..1123)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1446..1465
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 1640..1663
/label=DsRed1-C
/note="DsRed1, forward primer"
CDS 1732..1749
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1772..1789)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1778..2002
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2048..2476
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2135..2154)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2345..2366
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2485..2505)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2490..2819
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2670..2805
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2732..2751
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2867..2914
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2933..3331
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 3489..3610
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3526..3545)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3580..3599
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3659..3675)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3659..3675)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3672..3694)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3683..3699
/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(3707..3737)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3752..3773
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3890..3907)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4061..4649)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4141..4160)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4820..5680)
/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 5443..5462
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5681..5785)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttacca tggtggcctc ctccgaggac gtcatcaaag agttcatgcg
961 cttcaaggtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
1021 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
1081 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggcgtacgt
1141 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
1201 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
1261 gcaggacggc acgctgatct acaaggtgag tatggatccc tctaaaagcg ggcatgactt
1321 ctagagtagt ccagggtttc cgagggtttc cgtcgactca gctcgtctcg agggcgtact
1381 aactgggccc tttccctttt ttttcctcag gtgaagttcc gcggcaccaa cttccccccc
1441 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
1501 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
1561 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca gctgcccggc
1621 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
1681 gaacagtacg agcgctccga gggccgccac cacctgttcc tgtagaccgg tcatcatcac
1741 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
1801 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
1861 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
1921 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
1981 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
2041 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
2101 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
2161 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
2221 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
2281 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
2341 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
2401 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
2461 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
2521 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2641 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
2701 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
2761 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
2821 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
2881 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
2941 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
3001 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
3061 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
3121 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
3181 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
3241 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
3301 gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag caggactgac
3361 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
3421 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
3481 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
3541 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
3601 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
3661 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
3721 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
3781 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
3841 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
3901 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
3961 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
4021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
4081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
4141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
4261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
4321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
4381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
4441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
4501 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
4561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
4621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
4681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
4741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
4801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
4861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
4921 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
4981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
5041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
5101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
5161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
5401 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
5461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
5521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
5581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
5641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
5701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
5761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
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