| 质粒名称: | pBY2982 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 23220 |
| 存储实验室: | Ralf Baumeister |
|
载体骨架基本信息 |
|
| 载体名称: | EGFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 3992bp |
| 完整质粒大小: | 5702 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin |
| 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: | BirA-mCherry |
| 物种来源: | Top10 E.coli, Discosoma sp. |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pBY2982 was a gift from Ralf Baumeister (Addgene plasmid # 23220 ; http://n2t.net/addgene:23220 ; RRID:Addgene_23220) |
| 参考文献部分: | SnAvi - a new tandem tag for high-affinity protein-complex purification. Schaffer U, Schlosser A, Muller KM, Schafer A, Katava N, Baumeister R, Schulze E. Nucleic Acids Res. 2010 Jan 4. ():. 10.1093/nar/gkp1178 PubMed 20047968 |
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 pBY2982 5702 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBY2982
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5702)
AUTHORS Schaffer U, Schlosser A, Muller KM, Schafer A, Katava N, Baumeister
R, Schulze E
TITLE SnAvi - a new tandem tag for high-affinity protein-complex
purification.
JOURNAL Nucleic Acids Res. 2010 Jan 4. ():.
PUBMED 20047968
REFERENCE 2 (bases 1 to 5702)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5702
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 609..665
/label=MCS
/note="multiple cloning site"
CDS 667..1629
/codon_start=1
/gene="birA"
/product="E. coli biotin protein ligase"
/label=BirA
/translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
regulatory 1651..1660
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1657..2367
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1802..1820)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2070..2089
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2490..2611
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2527..2546)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2581..2600
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2618..3073)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2755..2776)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2967..2986
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3100..3204
/gene="bla"
/label=AmpR promoter
promoter 3206..3563
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3229..3249)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3414..3549
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3476..3495
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3598..4392
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3652..3671)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4262..4281
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4580..4599)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4624..4671
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5000..5588
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5489..5508
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gatccaatga aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa
721 tttcactctg gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa
781 cacattcaga cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac
841 agcctgcctg agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc
901 ggtagtgtag ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc
961 ggagagctta aatcgggcga tgcttgcatt gcagaatacc agcaggctgg ccgtggtcgc
1021 cggggtcgga aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt
1081 ctggaacaag gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg
1141 gaagtattac gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat
1201 ctgcaggatc gcaagctggc aggcattctg gtggagctga ctggcaaaac tggcgatgcg
1261 gcgcaaatag tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc
1321 gttaatcagg ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg
1381 gcggccatgc taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg
1441 gcaccttatc tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt
1501 atcattggtg ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta
1561 ttacttgagc aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt
1621 gcagaaaaaa ggagcatcgc cacaccggtc gccaccatgg tgagcaaggg cgaggaggat
1681 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
1741 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
1801 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
1861 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
1921 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
1981 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
2041 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
2101 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
2161 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
2221 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
2281 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
2341 ggcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
2401 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
2461 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
2521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
2581 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
2641 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
2701 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
2761 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
2821 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
2881 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
2941 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
3001 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
3061 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
3121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
3181 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
3241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
3481 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
3541 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
3601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
3661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
3721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
3781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
3841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
3901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
3961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
4021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
4081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
4141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
4201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
4261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
4321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
4381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
4441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
4501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
4561 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
4621 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
4681 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
4741 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
4801 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
4861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
4921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
4981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
5041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
5101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
5161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
5221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
5281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
5341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
5401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
5461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
5521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
5581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
5641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
5701 at
//
