| 质粒名称: | EJ2GFP-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44025 |
| 存储实验室: | Jeremy Stark |
|
载体骨架基本信息 |
|
| 载体名称: | pgkPURO |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EJ2GFP egfp-based chromosomal break reporter |
| 物种来源: | 合成 |
| 插入大小: | -- |
| 基因突变: | 插入I-SceI，3帧停止，侧翼微同源性 |
| GenBank编号 | U55761.1 |
| 启动子 | pCAGGS |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | ttcctacagctcctgggcaacg |
| 3' 测序引物 | ttttggcagagggaaaaaga |
|
文献引用方法 |
|
| 材料和方法部分: | EJ2GFP-puro was a gift from Jeremy Stark (Addgene plasmid # 44025 ; http://n2t.net/addgene:44025 ; RRID:Addgene_44025) |
| 参考文献部分: | Alternative-NHEJ is a mechanistically distinct pathway of mammalian chromosome break repair. Bennardo N, Cheng A, Huang N, Stark JM. PLoS Genet. 2008 Jun 27;4(6):e1000110. doi: 10.1371/journal.pgen.1000110. 10.1371/journal.pgen.1000110 PubMed 18584027 |
买家导航
EJ2GFPpuro载体基本信息
EJ2GFPpuro载体图谱质粒图谱和多克隆位点信息
EJ2GFPpuro载体简介
EJ2GFPpuro载体序列
LOCUS EJ2GFP-puro 7636 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EJ2GFP-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7636)
AUTHORS Bennardo N, Cheng A, Huang N, Stark JM
TITLE Alternative-NHEJ is a mechanistically distinct pathway of mammalian
chromosome break repair.
JOURNAL PLoS Genet. 2008 Jun 27;4(6):e1000110. doi:
10.1371/journal.pgen.1000110.
PUBMED 18584027
REFERENCE 2 (bases 1 to 7636)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7636
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 29..526
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(64..87)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 459..478
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 545..1144
/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(545..564)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1041..1061
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
enhancer 1788..2167
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2170..2445
/label=chicken beta-actin promoter
intron 2446..3457
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 3381..3403
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 3465..3484
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 3546..3566
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3588..3608
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 3857..3866
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3863..4582
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3908..3929)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4169..4188)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4516..4537
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(4787..4806)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4852..4907
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4906..4925)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
promoter complement(5263..5281)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5264..5281)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(5362..5381)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5481..5503
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5541..5559)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5627..5731
/gene="bla"
/label=AmpR promoter
CDS 5732..6592
/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 complement(5950..5969)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6763..7351
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7252..7271
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7505..7522
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 7609..7628
/label=T7
/note="T7 promoter, forward primer"
promoter 7609..7627
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 agatctgata tcatcgatga attctaccgg gtaggggagg cgcttttccc aaggcagtct
61 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
121 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
181 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
241 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
301 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
361 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
421 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
481 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgaccg atccagccgc
541 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
601 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
661 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
721 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
781 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
841 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
901 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
961 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
1021 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
1081 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
1141 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
1201 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgcg
1261 ggggacacac cgaacacgcc gaccctgctg aacacgcggc gcagttcggt gcccaggagc
1321 ggatcgaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga agtttttcct
1381 gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg aaggattgga
1441 gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact gaaggctctt
1501 tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca agcaaaacca
1561 aattaagggc cagctcattc ctcccactca tgatctatag atctatagat ctctcgtggg
1621 atcattgttt ttctcttgat tcccactttg tggttctaag tactgtggtt tccaaatgtg
1681 tcagtttcat agcctgaaga acgagatcag cagcctctgt tccacataca cttcattctc
1741 agtattgttt tgccaagttc taattccatc agaagcttca gctgctcgac attgattatt
1801 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
1861 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
1921 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
1981 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
2041 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
2101 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
2161 taccatgggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
2221 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
2281 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
2341 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
2401 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgttg
2461 ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct ctgactgacc
2521 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
2581 ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa agggctccgg
2641 gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
2701 gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
2761 ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
2821 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
2881 gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc cgagttgctg
2941 agcacggccc ggcttcgggt gcggggctcc gtgcggggcg tggcgcgggg ctcgccgtgc
3001 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
3061 agggctcggg ggaggggcgc ggcggccccg gagcgccggc ggctgtcgag gcgcggcgag
3121 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
3181 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
3241 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
3301 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
3361 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
3421 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
3481 tgtgctgtct catcattttg gcaaagaatt aattcagatc cgccgccact atgggatcaa
3541 gatcgccaaa aaagaagaga aaggtgccga agaagcatgc agcaccacca aaaaaaaaac
3601 gaaaagtaga agacccacga ggcaacacca gcggcgtgct gagcaccccc aaggccaaga
3661 gggccaagca cccccccggc accgagaagc ccaggagcag gagccagagc gagcagcccg
3721 ccacctgccc catctgctac gccgtgatca ggcagagcag gaacctgagg aggcacctgg
3781 agctgaggca cttcgccaag cccgcctagg gataacaggg taattagatg acaagcccgg
3841 cgtggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
3901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
3961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
4021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
4081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
4141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
4201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
4261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
4321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
4381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
4441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
4501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
4561 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
4621 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
4681 aaaatgaatg caattgttgt tgttaattca ctcctcaggt gcaggctgcc tatcagaagg
4741 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
4801 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
4861 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
4921 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
4981 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
5041 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
5101 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
5161 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
5221 ttatgaagat ccctcgacct gcagcccaag ctggcgtcga gttctatagt gtcacctaaa
5281 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat
5341 gtccatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
5401 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
5461 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
5521 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
5581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
5641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
5701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
5821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
5941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
6001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
6061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
6121 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
6181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
6241 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
6301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
6421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
6541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
6601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
6901 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
7141 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
7381 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
7441 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
7501 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
7561 cgcgcgttgg ccgattcatt aatgcaggtt aacctggctt atcgaaatta atacgactca
7621 ctatagggag accggc
//
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EJ2GFPpuro载体质粒应用举例
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