| 质粒名称: | pCMV/SB10 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24551 |
| 存储实验室: | Perry Hackett |
|
载体骨架基本信息 |
|
| 载体名称: | pCMVbeta |
| 载体出品公司: | Clontech |
| 空载体大小: | 3712bp |
| 完整质粒大小: | 4732bp |
| 载体类型: | 细菌表达 |
| 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: | Sleeping Beauty、SB、SB10 |
| 插入大小: | 1020bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI (destroyed during cloning) |
| 3’克隆位点: | NotI (destroyed during cloning) |
| 5’测序引物 | M13 Forward |
| 3’测序引物 | M13 Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV/SB10 was a gift from Perry Hackett (Addgene plasmid # 24551 ; http://n2t.net/addgene:24551 ; RRID:Addgene_24551) |
| 参考文献部分: | Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Ivics Z, Hackett PB, Plasterk RH, Izsvak Z. Cell. 1997 Nov 14. 91(4):501-10. 10.1016/S0092-8674(00)80436-5 PubMed 9390559 |
买家导航
pCMV/SB10载体质粒基本信息
pCMV/SB10质粒图谱载体图谱和pCMV/SB10载体序列质粒序列多克隆位点信息
pCMV/SB10质粒载体简介
pCMV/SB10质粒序列载体序列
LOCUS pCMV/SB10 4732 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4732)
AUTHORS Ivics Z, Hackett PB, Plasterk RH, Izsvak Z
TITLE Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon
from fish, and its transposition in human cells.
JOURNAL Cell. 1997 Nov 14. 91(4):501-10.
PUBMED 9390559
REFERENCE 2 (bases 1 to 4732)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1997
Nov 14. 91(4):501-10."
FEATURES Location/Qualifiers
source 1..4732
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(23..40)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(194..782)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(274..293)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(953..1813)
/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 1576..1595
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1814..1918)
/gene="bla"
/label=AmpR promoter
primer_bind 1986..2004
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2042..2064)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2164..2183
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2377..2399
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2391..2408
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2392..2408
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 2435..2738
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2739..2942
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2893..2913
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2939..2963
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 3080..3176
/label=SV40 intron
/note="modified SV40 intron with splice donor and acceptor
sites"
CDS 3243..4265
/codon_start=1
/product="hyperactive variant of the Sleeping Beauty
transposase (Mates et al., 2009)"
/label=SB100X
/translation="MGKSKEISQDLRKKIVDLHKSGSSLGAISKRLKVPRSSVQTIVRK
YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
RVLYRHNLKGRSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMRKENYVDIL
KQHLKTSVRKLKLGRKWVFQMDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPTYCGKLVEGYPKRLTQVKQFKGNATKY
"
polyA_signal 4279..4413
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4303..4322)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4357..4376
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4524..4540)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4524..4540)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4537..4559)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4548..4564
/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(4572..4602)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4617..4638
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
1921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
1981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
2041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
2101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
2161 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
2221 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
2281 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
2341 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
2401 cggccagtga attcgagctt gcatgcctgc aggtcgttac ataacttacg gtaaatggcc
2461 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
2521 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
2581 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
2641 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
2701 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
2761 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
2821 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
2881 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
2941 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
3001 gaagacaccg ggaccgatcc agcctccgga ctctagagga tccggtactc gaggaactga
3061 aaaaccagaa agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc
3121 cggtggtggt gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct
3181 gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgatccgaca
3241 tcatgggaaa atcaaaagaa atcagccaag acctcagaaa aaaaattgta gacctccaca
3301 agtctggttc atccttggga gcaatttcca aacgcctgaa agtaccacgt tcatctgtac
3361 aaacaatagt acgcaagtat aaacaccatg ggaccacgca gccgtcatac cgctcaggaa
3421 ggagacgcgt tctgtctcct agagatgaac gtactttggt gcgaaaagtg caaatcaatc
3481 ccagaacaac agcaaaggac cttgtgaaga tgctggagga aacaggtaca aaagtatcta
3541 tatccacagt aaaacgagtc ctatatcgac ataacctgaa aggccgctca gcaaggaaga
3601 agccactgct ccaaaaccga cataagaaag ccagactacg gtttgcaact gcacatgggg
3661 acaaagatcg tactttttgg agaaatgtcc tctggtctga tgaaacaaaa atagaactgt
3721 ttggccataa tgaccatcgt tatgtttgga ggaagaaggg ggaggcttgc aagccgaaga
3781 acaccatccc aaccgtgaag cacgggggtg gcagcatcat gttgtggggg tgctttgctg
3841 caggagggac tggtgcactt cacaaaatag atggcatcat gaggaaggaa aattatgtgg
3901 atatattgaa gcaacatctc aagacatcag tcaggaagtt aaagcttggt cgcaaatggg
3961 tcttccaaat ggacaatgac cccaagcata cttccaaagt tgtggcaaaa tggcttaagg
4021 acaacaaagt caaggtattg gagtggccat cacaaagccc tgacctcaat cctatagaaa
4081 atttgtgggc agaactgaaa aagcgtgtgc gagcaaggag gcctacaaac ctgactcagt
4141 tacaccagct ctgtcaggag gaatgggcca aaattcaccc aacttattgt gggaagcttg
4201 tggaaggcta cccgaaacgt ttgacccaag ttaaacaatt taaaggcaat gctaccaaat
4261 actagaattg gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc
4321 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
4381 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
4441 tttcaggttc agggggaggt gtgggaggtt ttttcggatc ctctagagtc gacctgcagg
4501 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
4561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
4621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
4681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gc
//
