| 质粒名称: | CMV-Tol2 transposase |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 158774 |
| 存储实验室: | Hugo Snippert |
|
载体骨架基本信息 |
|
| 载体名称: | pDB739 |
| 载体出品公司: | Clontech |
| 空载体大小: | 2927bp |
| 完整质粒大小: | 6108bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Tol2 tranposase |
| 物种来源: | Synthetic |
| 插入大小: | 1950bp |
| 基因突变: | Tol2 codon usage frequency well suitable for murine or human cells |
|
文献引用方法 |
|
| 材料和方法部分: | CMV-Tol2 transposase was a gift from Hugo Snippert (Addgene plasmid # 158774 ; http://n2t.net/addgene:158774 ; RRID:Addgene_158774) |
| 参考文献部分: | Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal Organoids. Heinz MC, Oost KC, Snippert HJG. STAR Protoc. 2020 Oct 8;1(3):100126. doi: 10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18. 10.1016/j.xpro.2020.100126 PubMed 33377020 |
买家导航
CMV-Tol2 transposase载体质粒基本信息
CMV-Tol2 transposase质粒图谱载体图谱和CMV-Tol2 transposase载体序列质粒序列多克隆位点信息
CMV-Tol2 transposase质粒载体简介
CMV-Tol2 transposase质粒序列载体序列
LOCUS CMV-Tol2 transposase 6018 bp DNA circular SYN 15-DEC-2025
DEFINITION Expression of Tol2 transposase.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6018)
AUTHORS Heinz MC, Oost KC, Snippert HJG
TITLE Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal
Organoids.
JOURNAL STAR Protoc. 2020 Oct 8;1(3):100126. doi:
10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18.
PUBMED 33377020
REFERENCE 2 (bases 1 to 6018)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofenge.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "STAR
Protoc."; date: "2020-10-8"; volume: "1(3):100126. doi"; pages: "
10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18"
COMMENT Sequence Label: CMV-Tol2 transposase
FEATURES Location/Qualifiers
source 1..6018
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..978
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1013..1031
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1013..1030
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 1063..1072
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1069..3018
/codon_start=1
/product="transposase of the Tol2 transposon from the
medaka fish Oryzias latipes (Kawakami, 2007)"
/label=Tol2 transposase
/translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
QLLLKLNLRFYNFE"
polyA_signal 3048..3182
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3072..3091)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3126..3145
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3295..3315)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3295..3313)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3334..3350)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3334..3350)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3347..3369)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3358..3374
/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(join(3382..3388,3389..3406,3407..3412))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3427..3448
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3565..3582)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3736..4324)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3816..3835)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4495..5286,5287..5355))
/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 5118..5137
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5356..5460)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5486..5941)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5623..5644)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5835..5854
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
1021 gacactatag aatacaagct acttgttctt tttgcaggat ccgccaccat ggaagaggtg
1081 tgcgactctt ccgccgcagc ttcatctacc gtccagaacc agcctcagga ccaagagcac
1141 ccctggccat atctgagaga gttcttctcc ctgagcggag tgaataagga ctcatttaaa
1201 atgaagtgtg tgctgtgtct tccactcaac aaagagatta gcgcgtttaa atcctcaccc
1261 tcgaacctga ggaagcacat tgagcgtatg caccctaact acctgaaaaa ttactccaag
1321 ctgaccgccc agaagagaaa gataggcact agtactcacg catccagcag taagcagttg
1381 aaagtagact cagtgttccc tgttaagcat gtgtcccctg taacagttaa caaggctatt
1441 ctccggtaca ttatccaagg tctgcacccg ttcagcaccg tcgacctacc ctccttcaag
1501 gagctgatta gtacgcttca gcctggaatt agtgtgatta ctcgccctac tctgagaagc
1561 aaaatcgcag aagcggcgct gatcatgaaa cagaaagtga ctgctgcaat gagcgaggtg
1621 gaatggatcg cgacgacaac cgactgttgg acagcacgta ggaagtcctt tataggcgtc
1681 actgcccact ggattaaccc cggttcactg gagcgacact cagcagccct ggcctgcaaa
1741 cgccttatgg gatctcacac ttttgaggtg ttagcgtctg ctatgaacga catccactcg
1801 gaatacgaaa ttagggacaa ggtggtctgt accacaacag actctggatc taactttatg
1861 aaagcgtttc gcgtttttgg cgtggagaac aatgatatcg aaacggaggc cagacgctgt
1921 gagtcagacg acactgacag cgaaggctgc ggcgaaggtt cagacggcgt cgaatttcag
1981 gacgcgagtc gcgttctcga ccaggatgat ggatttgaat ttcagttacc aaagcaccag
2041 aagtgtgcct gtcatcttct aaacctcgtc tcttcggtcg acgcacaaaa agctctgtcc
2101 aacgagcatt acaagaagct ctaccggtct gtcttcggca agtgtcaggc cctttggaac
2161 aagtcctcca gatccgctct cgccgcagag gcggtagaga gtgagtccag acttcagctg
2221 ctgagaccta atcagacacg ctggaacagc actttcatgg cagtggaccg aatcttacaa
2281 atctgtaaag aagccggtga gggcgctctc cgcaacatct gtacatcact ggaagtccct
2341 atgtttaacc cagctgagat gctcttcttg acagagtggg ctaacactat gcggccagtc
2401 gctaaagtcc tcgacatcct tcaggccgaa accaatacac agttaggctg gctccttcca
2461 agcgtgcacc aactcagtct gaagctccaa agactgcatc atagtctgcg gtactgcgat
2521 cccctcgtgg atgctctgca gcagggaatc cagactcggt tcaaacacat gttcgaagac
2581 cctgagatta tcgctgcagc catccttctc ccaaaatttc gaacctcgtg gacaaacgat
2641 gagaccatta ttaagcgcgg catggattat atccgcgtcc accttgaacc cctggaccac
2701 aaaaaagagc tggctaattc aagtagtgac gatgaggatt tctttgccag cttgaagccg
2761 acgacacacg aggcctcaaa ggagctggac ggttacctgg cctgcgtcag cgatacaagg
2821 gagtcactgt tgaccttccc tgcaatctgt tcgctgtcga tcaagacaaa cactcccttg
2881 cctgctagtg cagcgtgtga gagactcttt agcacagcag gactcctgtt cagccctaag
2941 agagcaaggc ttgataccaa caactttgag aaccagttgc ttctgaaact gaatttacga
3001 ttttacaact tcgagtgatc tagaactata gtgagtcgta ttacgtagat ccagacatga
3061 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
3121 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
3181 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
3241 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
3301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
3361 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
3421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
3481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
3541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
3601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
4141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
4501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
4561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
4621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
4681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
4741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
4801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
4861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
4921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
4981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
5041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
5101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
5161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
5221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
5281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
5341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
5401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
5461 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
5521 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
5581 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
5641 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
5701 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
5761 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
5821 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
5881 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
5941 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
6001 ctattacgcc agtcgacc
//
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