| 质粒名称: | pcDNA3.1 puro Nodamura B2 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 17228 |
| 存储实验室: | Christopher Sullivan |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 puro+ |
| 空载体大小: | 5500 bp |
| 完整质粒大小: | 5531bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 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: | Nodamura B2 |
| 物种来源: | Nodamura Virus |
| 插入大小: | 429 bp |
| 基因突变: | 2 silent point mutations |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI(未破坏) |
| 3’克隆位点: | XbaI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: |
pcDNA3.1 puro Nodamura B2 was a gift from Christopher Sullivan (Addgene plasmid # 17228 ; http://n2t.net/addgene:17228 ; RRID:Addgene_17228)
|
| 参考文献部分: | A virus-encoded inhibitor that blocks RNA interference in mammalian cells. Sullivan CS, Ganem D. J Virol. 2005 Jun . 79(12):7371-9. 10.1128/JVI.79.12.7371-7379.2005 PubMed 15919892 |
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 pcDNA3.1 puro Nodamura B2 5531 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 puro Nodamura B2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5531)
AUTHORS Sullivan CS, Ganem D
TITLE A virus-encoded inhibitor that blocks RNA interference in mammalian
cells.
JOURNAL J Virol. 2005 Jun . 79(12):7371-9.
PUBMED 15919892
REFERENCE 2 (bases 1 to 5531)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5531
/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"
primer_bind complement(1377..1394)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1383..1607
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1653..2081
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1740..1759)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1950..1971
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2090..2110)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2095..2424
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2275..2410
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2337..2356
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2445..3044
/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(2445..2464)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2941..2961
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 3204..3325
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3241..3260)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3295..3314
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3374..3390)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3374..3390)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3387..3409)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3398..3414
/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(3422..3452)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3467..3488
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3605..3622)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3776..4364)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3856..3875)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4535..5395)
/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 5158..5177
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5396..5500)
/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 aagcttggta ccgccgccac catgacaaac atgtcatgcg cttacgagct
961 aatcaagtca cttccagcca agctggagca gctggctcag gagacgcaag caacgatcca
1021 aacgctcatg atcgcggatc ccaacgtcaa caaggatctg cgagcgttct gcgagttcct
1081 gaccgtacag caccagcggg cgtatcgagc gacgaacagc ctgctcatca aaccgcgagt
1141 cgcagcagcg cttcgcgggg aggagctgga cctgggcgag gcggacgtcg ccgcccgggt
1201 ccgccagcta aaacaacagc tggcggagct cgagatggaa atcaagccag ggcaccaaca
1261 ggtggcccaa gtaagcggca ggcggaaggc cgcagccgca gctcccgtgg cccagctggg
1321 tcgcgtgggc gtggtaaatg agtgatctag agggcccgtt taaacccgct gatcagcctc
1381 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
1441 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
1501 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
1561 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
1621 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
1681 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
1741 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
1801 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
1861 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
1921 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
1981 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
2041 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
2101 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2161 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2221 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
2281 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
2341 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
2401 tttggaggcc taggcttttg caaaaagctc ccggggccgc caccatgacc gagtacaagc
2461 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
2521 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
2581 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
2641 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
2701 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
2761 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
2821 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
2881 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
2941 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
3001 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgattcgaa atgaccgacc
3061 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
3121 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
3181 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
3241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
3301 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
3361 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
3421 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
3481 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
3541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
3601 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
3661 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
3721 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
3781 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
3841 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
3901 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
3961 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
4021 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
4081 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
4141 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
4201 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
4261 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
4321 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
4381 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
4441 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
4501 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
4561 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
4621 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
4681 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
4741 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
4801 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
4861 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
4921 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
4981 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
5041 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
5101 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
5161 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
5221 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
5281 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
5341 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
5401 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
5461 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
5521 cacctgacgt c
//
