| 质粒名称: | pUGI-NLS |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101091 |
| 存储实验室: | Jia Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 完整质粒大小: | 5714 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | UGI / UGI-NLS |
| 物种来源: | Bacteriophage PBS2 |
| 插入大小: | 276 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | NLS (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | known (unknown if destroyed) |
| 3’克隆位点: | known (unknown if destroyed) |
| 5’测序引物 | CMV |
| 3’测序引物 | BGH poly reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pUGI-NLS was a gift from Jia Chen (Addgene plasmid # 101091 ; http://n2t.net/addgene:101091 ; RRID:Addgene_101091) |
| 参考文献部分: | Enhanced base editing by co-expression of free uracil DNA glycosylase inhibitor. Wang L, Xue W, Yan L, Li X, Wei J, Chen M, Wu J, Yang B, Yang L, Chen J. Cell Res. 2017 Aug 29. doi: 10.1038/cr.2017.111. 10.1038/cr.2017.111 PubMed 28849781 |
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 pUGI-NLS 5714 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses UGI in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pUGI-NLS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5714)
AUTHORS Wang L, Xue W, Yan L, Li X, Wei J, Chen M, Wu J, Yang B, Yang L,
Chen J
TITLE Enhanced base editing by co-expression of free uracil DNA
glycosylase inhibitor.
JOURNAL Cell Res. 2017 Aug 29. doi: 10.1038/cr.2017.111.
PUBMED 28849781
REFERENCE 2 (bases 1 to 5714)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5714
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 41..292
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 293..313
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(361..378)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 367..591
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 637..1065
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(724..743)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 934..955
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1074..1094)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1079..1408
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1259..1394
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1321..1340
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1627..1646
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 1715..2314
/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(1715..1734)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2211..2231
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 2488..2609
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2525..2544)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2579..2598
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2658..2674)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2658..2674)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2671..2693)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2682..2698
/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(2706..2736)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2751..2772
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2889..2906)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3060..3648)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3140..3159)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3819..4679)
/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 4442..4461
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4680..4784)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4859..4878)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 5050..5429
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5430..5633
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5584..5604
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5678..5697
/label=T7
/note="T7 promoter, forward primer"
promoter 5678..5696
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 actcgagcgg ccgccactgt gctggatatc tgcagaattc atgactaatc tgtcagatat
61 tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga
121 ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta
181 cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc
241 ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc tccccaagaa
301 gaagaggaaa gtctagggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag
361 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
421 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
481 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
541 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
601 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
661 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
721 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
781 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
841 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
901 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
961 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
1021 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
1081 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
1141 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
1201 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
1261 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
1321 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
1381 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
1441 atctgatcaa gagacaggat gaggatcgtt tcgcatggaa gcgggtaggc ctttggggca
1501 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
1561 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
1621 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
1681 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
1741 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
1801 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
1861 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
1921 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
1981 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
2041 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
2101 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
2161 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
2221 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
2281 cacctggtgc atgacccgca agcccggtgc ctgagcggga ctctggggtt cgaaatgacc
2341 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
2401 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
2461 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
2521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
2581 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
2641 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
2701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
2761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
2821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
2881 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
2941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
3001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
3061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
3121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
3181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
3241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
3301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
3361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
3421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
3481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
3541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
3601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
3661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
3721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
3781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
3841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
3901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
3961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
4021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
4081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
4141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
4201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
4261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
4321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
4381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
4441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
4501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
4561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
4621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
4681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
4741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
4801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
4861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
4921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
4981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
5041 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
5101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
5161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
5221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
5281 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
5341 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
5401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
5461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
5521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
5581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
5641 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
5701 cccaagctgg ctag
//
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