| 质粒名称: | pWZL hygro 12S E1A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 18748 |
| 存储实验室: | Scott Lowe |
|
载体骨架基本信息 |
|
| 载体名称: | pWZL Hygro |
| 载体出品公司: | Available at Addgene (#18750) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5915 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 潮霉素 |
| 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: | 12S E1A,E1A 12S,E1a protein from 12s mrna |
| 物种来源: | adenovirus |
| 插入大小: | 701 bp |
| 基因突变: | -- |
| GenBank编号 | CAB40664.1 |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI (未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | pBABE 5' |
| 3' 测序引物 | IRES-R |
|
文献引用方法 |
|
| 材料和方法部分: | pWZL hygro 12S E1A was a gift from Scott Lowe (Addgene plasmid # 18748 ; http://n2t.net/addgene:18748 ; RRID:Addgene_18748) |
| 参考文献部分: | Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW. Cell. 1997 Mar 7. 88(5):593-602. 10.1016/S0092-8674(00)81902-9 PubMed 9054499 |
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 pWZL hygro 12S E1A 6437 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pWZL hygro 12S E1A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6437)
AUTHORS Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW
TITLE Oncogenic ras provokes premature cell senescence associated with
accumulation of p53 and p16INK4a.
JOURNAL Cell. 1997 Mar 7. 88(5):593-602.
PUBMED 9054499
REFERENCE 2 (bases 1 to 6437)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6437
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1288..1310
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1326..1342
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 2251..2712
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2307..2324)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2534..2553
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2717..3742
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
LTR 3931..4400
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4530..4547)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4701..5289)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4781..4800)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5449..6309)
/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 6072..6091
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6310..6414)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
1381 acgtaggaat tcaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg
1441 gccgccagtc ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct
1501 agccattttg aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa
1561 gatcccaacg aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag
1621 gaagggattg acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt
1681 tcccggcagc ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt
1741 gtaccggagg tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag
1801 gatgaagagg gtcctgtgtc tgaacctgag cctgagcccg agccagaacc ggagcctgca
1861 agacctaccc gccgtcctaa aatggcgcct gctatcctga gacgcccgac atcacctgtg
1921 tctagagaat gcaatagtag tacggatagc tgtgactccg gtccttctaa cacacctcct
1981 gagatacacc cggtggtccc gctgtgcccc attaaaccag ttgccgtgag agttggtggg
2041 cgtcgccagg ctgtggaatg tatcgaggac ttgcttaacg agcctgggca acctttggac
2101 ttgagctgta aacgccccag gccataaggt gtaaacctgt gattgcgtgt gtggttcggg
2161 atcccagtgt ggtggtacgt aggaattcgc cagcacagtg gtcgaccaat tccggttatt
2221 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
2281 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
2341 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
2401 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
2461 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
2521 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
2581 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
2641 gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
2701 acacgatgat aataccatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct
2761 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
2821 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
2881 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
2941 ggaagtgctt gacattgggg aatttagcga gagcctgacc tattgcatct cccgccgtgc
3001 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
3061 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
3121 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
3181 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
3241 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
3301 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
3361 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
3421 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
3481 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
3541 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
3601 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
3661 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
3721 tcgtccgagg gcaaaggaat agagtagatg ccgaccggga tctatcgata aaataaaaga
3781 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
3841 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
3901 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
3961 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
4021 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
4081 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
4141 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
4201 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
4261 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
4321 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
4381 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
4441 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
4501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
5101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
5281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
5341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
5401 cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
5461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
5521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
5581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
5641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
5701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
5761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
5821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
5881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
5941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
6001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
6061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
6121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
6181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
6241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
6301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
6361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
6421 tccccgaaaa gtgccac
//
