| 质粒名称: | pDestTol2pA2_ubi:EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 27323 |
| 存储实验室: | Leonard Zon |
|
载体骨架基本信息 |
|
| 载体名称: | pDestTol2A2 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4179 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Tol2转座子 |
| 筛选标记: | -- |
| 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: | ubiquitin:EGFP reporter,ubi:EGFP,ubi-EGFP,ubi-GFP |
| 物种来源: | D. rerio (zebrafish) |
| 插入大小: | 4606 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | attB4(未破坏) |
| 3' 克隆位点: | attB3 (未破坏) |
| 5' 测序引物 | attB4 |
| 3' 测序引物 | attB3 |
|
文献引用方法 |
|
| 材料和方法部分: | pDestTol2pA2_ubi:EGFP was a gift from Leonard Zon (Addgene plasmid # 27323 ; http://n2t.net/addgene:27323 ; RRID:Addgene_27323) |
| 参考文献部分: | Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI. Development. 2011 Jan . 138(1):169-77. 10.1242/dev.059345 PubMed 21138979 |
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 pDestTol2pA2_ubi:EGFP 8786 bp ds-DNA circular SYN 08-NOV-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDestTol2pA2_ubi:EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8786)
AUTHORS Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI
TITLE Ubiquitous transgene expression and Cre-based recombination driven
by the ubiquitin promoter in zebrafish.
JOURNAL Development. 2011 Jan . 138(1):169-77.
PUBMED 21138979
REFERENCE 2 (bases 1 to 8786)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 8, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8786
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(21..37)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 53..187
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(859..875)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(859..875)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(912..932)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(912..930)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(951..967)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(964..986)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 975..991
/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(999..1029)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1044..1065
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1182..1199)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1353..1941)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1433..1452)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2112..2972)
/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 2735..2754
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2973..3077)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3240..3261)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4157..4173
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4193..4213
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 7748..7772
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 7778..8494
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7823..7844)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(8084..8103)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8431..8452
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(8556..8580)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 8582..8716
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(8779..13)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
ORIGIN
1 ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga
61 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
121 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
181 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
241 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
301 tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg
361 ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt
421 ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac
481 gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt
541 tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt
601 ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt
661 aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa
721 gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt
781 aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta
841 ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
901 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
1021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
1081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
1141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
1201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
1261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
1321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
1381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
1441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
1501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
1561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
1621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
1681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
1741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
1801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
1861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
1921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
1981 tggaacgaaa actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc
2041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
2101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
2161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
2221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
2281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
2341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
2401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
2461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
2521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
2581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
2641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
2701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
2761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
2821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
2881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
2941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
3001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
3061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
3121 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
3181 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
3241 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
3301 ccgtctatca gggcgatggc ccacgctggc taagaactca tcagcctccc cggtccatct
3361 acccacgtac caatgcacca attggccaca atgacggcta ctacatggtg ccattccttc
3421 ctctttatag gaatggagac tacctcctgt ccaacaatgc tcttggatac gagtacgcct
3481 acctgttgga cccaggtcat tgcacaacac cagaaatgcc ctctgatctg caaaagacgt
3541 gaatatctgt tcagacaccc atatccactc tgttccacac aggtcagagg tttgtccagg
3601 agttcttgac agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac
3661 ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa
3721 aagtaaaaaa gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca
3781 agaaagaaaa ctagagattc ttgtttaagc ttttaatctc aaaaaacatt aaatgaaatg
3841 catacaaggt tttatcctgc tttagaactg tttgtattta attatcaaac tataagacag
3901 acaatctaat gccagtacac gctactcaaa gttgtaaaac ctcagattta acttcagtag
3961 aagctgattc tcaaaattgt tagtgtcaag cctagctctt ttggggctga aaagcaatcc
4021 tgcagtgctg aaaagcctct cacaggcagc cgatgcggga agaggtgtat tagtcttgat
4081 agagaggctg caaatagcag gaaacgtgag cagagactcc ctggtgtctg aaacacaggc
4141 cagatgggcc ctcgagcagg aaacagctat gaccatgatt acgccaagct atcaactttg
4201 tatagaaaag ttggctccga attcgccctt aaaactcgag accagcaaag ttctagaatt
4261 tgtcgaaaca tttatgttat atatttcctg aaaaaaattc tgagtaagtt cttaagtgtt
4321 attgccagca acataaacaa cagacggcaa aatgaataaa tgataacaaa gcagtaggct
4381 taaataaacc taatttttat aggctgttct ctacaaccct caaacagtga ttagttttgt
4441 acttataaac ttgccctttc attcatattt caagaaaatt ggttcagaag atctggatat
4501 tctagcagtt gttcaagctc atggagggat cagtgacctg attccacaat gactaggcct
4561 aatccagaaa ttagatgact gtcaacataa aaaggcacag cactcactag ctgccctata
4621 tattttatta tattttacat atattatttt atttatttag ctctgagtgc tgtactttct
4681 ggttaaagaa aactgcttac aacagctaac ctgtactacc tcaggctcag ggaatttgga
4741 acaggtttgt ctggtttgtt tctttaacca tgcatgcttg ttttcaacta tggcaacaca
4801 gtcacatggg acattacaga aatgatttgt cgatgacatg cgacttttct ttaataaagc
4861 gcaaagatcc caaaaagcaa acttttaaca aaaatcatat aattatattt tcaatccagc
4921 tttgtagcaa ctttgtgctg ctgttcactc agcaacagat agtcagtata aggtcagtgt
4981 gtctcaaagc agtgccatct gtttcacaca ttgcgttcta tatataagtg tgctggttga
5041 cacgacactg tataaggcct aggctaaaac acaaacaatg tagaatgaca ctgtgttttt
5101 tttgtaaaca aatgttgttt ttggttaaac atctttgtga aaacatcctc ctgtcatgta
5161 tttgctatat tcaaatgtta aacccgtgca gaatagaaca tatacaaaaa aaaacaacac
5221 aacacatttt taaacattat taaatatcaa gtattgctgg cagttctgtt tctgttttac
5281 agtacccttt gccacagttc tccgcttttc ctggtccaga ttccacaagt ctgattcacc
5341 aatagcaaag cgaataaaca accaaagcag ccaatcactg cttgtagact gtcctgcgag
5401 accggcccat tccagcacat tctggaaact tcctttatat gataattata aatacattta
5461 aattattgat acaaaacatg taattcctag aacataacca tagcaatcat tagttttcag
5521 ggtaattatg tatttttagg atttgactgc ggaaagatct ggtcatgtga cgtctcatga
5581 acgtcacggc cctgggtttc tataaataca gtaggactct cgaccatcgg cagatttttc
5641 gaagaagaag atcagtttca ggagccgtac tgttccgttt caacgcaaat attaacggta
5701 agagcgaatt tcctagtttg ttttcatgcc attctttaaa accatagcgt attactttaa
5761 ttatagtaaa ctttcgcttt ctttattaca agagacgttt tgtgttgatt ctccgcggac
5821 attttcggtc agacaatcag aaaatgaccg cggaggacca gtaacttgca ttacacgtaa
5881 gttaaatctt cgtgtattaa aatggttagg ttgttaacgt caaataggtt accgtgtttg
5941 cgtgtgatca ggttggtttt gttagatttt tgtcagtatt tttaatttat ttgttttagt
6001 ttatttattt tttttgctga atcatagttt gtgaacaaag aacccggatg ttacatacag
6061 tacagccgcc atgttacaga gagttataac ttaatcattt taaaaataat tttgccttac
6121 ttttagtttg tcatgttgag aaatgaggaa atgttaaaat gaggaaatat ccaattaatt
6181 taatatatca aaataatcca tgattacaat gcactgaact ggagaaaatt aagatgtttt
6241 ctagtgtcat gaaacaaatg taagagatgt acattgtaga tgttttatgt caagaattgg
6301 ctagttgatg cagcatactg gcgatactca gttgtaataa cagtaacgtt acatgttaat
6361 agactactga gtatgctgtt ctgtctatgt atgctctgta agctacgaga aggacttttt
6421 taaacagtaa agggtgcaat atttttacaa attgaattaa ataaaggctg tctattaagt
6481 aatatgcttg atatttttct tacttgatcg gaaataagaa aaaatataaa cgttgttgct
6541 ctaaaaatcc tagttcagtt tagccaacca caaatacctt tttgttcctc caacagtttt
6601 tttttcttct ctataatatt tggcagtcta tagtactcca aatgtttccc cacagtctaa
6661 ctaattggta cagccaaaat catgacactt attgcaataa taattttggt tcattggcat
6721 tgttgatagc ctgtgccact aatatggtcg attgatcatg cttcaggaag aaaactatat
6781 tgtttgatgt aagattatta aatcttcacc tgcttccatt acaaactatt cccatcttat
6841 tgaattctgg tatgtcttaa aggattagtt cacttcccaa atcaaaattt acttagcctt
6901 ttttcatcca tgatcccttt ttttcatcat taatgaagaa attgtttttg aaaaagtttc
6961 aagatttttt tctctatatt gtggagcttg ttagtttaaa attccaaaat gcaatatgtg
7021 gcttcaaatg gttctaaatg atcccagtca aggaataaca gtcttatcta atgaaaccat
7081 tagaccttta aaaataaaaa aaataaaagt atttattttt aaatgactga gtgattaagt
7141 tgaatttcag cgtttcctta ctgtgtagaa gtccttcctt actggcccca ccctttggtt
7201 ctctgccaat ctgctaccta atgtaatgtt gtggaacatt attattcttt atttcttaat
7261 tttttatttt ttattttaaa acaatgtaaa ctgcacagat gtgcagtttg tttaaaatgg
7321 ccaatgcttt ggaaatgcat gacataatta gatttcatga tgcacaaagc caaatctcag
7381 agcttgtgca aaatgagcta tcatttcact aggtaagacc ctaaattttc atataggatc
7441 atttggacaa ttttgctgca ggtaaaatgc attctatagt ccactgtcag ccattgtttt
7501 ggatagtatt tatttttctc tacaagtata gtcaatagtt ttctattatt ttaaaggttt
7561 gtaacattta agggtgacca aatgcaaagt aaaatttcat tttcgggtga actatctcgt
7621 ttaacatggg agaagtgcaa aacatacatt attggctaga acattgtagt attttttaaa
7681 tggaaatgtg tgattgctaa tcttactttg aatttgttta cagggatcct ttaagggcga
7741 attcgaccca agtttgtaca aaaaagcagg ctggaccatg gtgagcaagg gcgaggagct
7801 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
7861 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
7921 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
7981 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
8041 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
8101 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
8161 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
8221 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
8281 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
8341 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
8401 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
8461 cgggatcact ctcggcatgg acgagctgta caagggcggt ggaagatctg ggaattcaag
8521 gcctctcgag cctctagatt ctgcagccct atagcaccca gctttcttgt acaaagtggg
8581 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
8641 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
8701 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
8761 ggtgtgggag gttttttcca acttta
//
