| 质粒名称: | pENTR5'_ubi:loxP-EGFP-loxP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 27322 |
| 存储实验室: | Leonard Zon |
|
载体骨架基本信息 |
|
| 载体名称: | pENTR5' TOPO |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 2692 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 多站点网关5'入口向量 |
| 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:loxP-EGFP-loxP,ubi:Switch,ubi:loxP-EGFP-loxP |
| 物种来源: | D. rerio (zebrafish) |
| 插入大小: | 5855 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | attL4(未破坏) |
| 3' 克隆位点: | attR1(未破坏) |
| 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: | pENTR5'_ubi:loxP-EGFP-loxP was a gift from Leonard Zon (Addgene plasmid # 27322 ; http://n2t.net/addgene:27322 ; RRID:Addgene_27322) |
| 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 |
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载体基本信息
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载体序列
LOCUS pENTR5'_ubi:loxP-EGFP-loxP 7496 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pENTR5'_ubi:loxP-EGFP-loxP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7496)
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 7496)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7496
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator 268..295
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 593..688
/gene="mutant version of attL"
/label=attL4
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 4210..4243
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
regulatory 4250..4259
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4256..4975
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4301..4322)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4562..4581)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4909..4930
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 5098..5219
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5135..5154)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5189..5208
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5363..5384)
/label=F1ori-F
/note="F1 origin, forward primer"
protein_bind 5473..5506
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 5537..5661
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(5737..5756)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5738..5756)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5761..5777)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5761..5777)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5879..5898)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 5890..6699
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(5957..5976)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 6846..7434
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7335..7354
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccga gttaacgcta ccatggagct ccaaataatg
601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
661 ataatgccaa ctttgtatag aaaagttggc tccgaattcg cccttaaact cgagaccagc
721 aaagttctag aatttgtcga aacatttatg ttatatattt cctgaaaaaa attctgagta
781 agttcttaag tgttattgcc agcaacataa acaacagacg gcaaaatgaa taaatgataa
841 caaagcagta ggcttaaata aacctaattt ttataggctg ttctctacaa ccctcaaaca
901 gtgattagtt ttgtacttat aaacttgccc tttcattcat atttcaagaa aattggttca
961 gaagatctgg atattctagc agttgttcaa gctcatggag ggatcagtga cctgattcca
1021 caatgactag gcctaatcca gaaattagat gactgtcaac ataaaaaggc acagcactca
1081 ctagctgccc tatatatttt attatatttt acatatatta ttttatttat ttagctctga
1141 gtgctgtact ttctggttaa agaaaactgc ttacaacagc taacctgtac tacctcaggc
1201 tcagggaatt tggaacaggt ttgtctggtt tgtttcttta accatgcatg cttgttttca
1261 actatggcaa cacagtcaca tgggacatta cagaaatgat ttgtcgatga catgcgactt
1321 ttctttaata aagcgcaaag atcccaaaaa gcaaactttt aacaaaaatc atataattat
1381 attttcaatc cagctttgta gcaactttgt gctgctgttc actcagcaac agatagtcag
1441 tataaggtca gtgtgtctca aagcagtgcc atctgtttca cacattgcgt tctatatata
1501 agtgtgctgg ttgacacgac actgtataag gcctaggcta aaacacaaac aatgtagaat
1561 gacactgtgt tttttttgta aacaaatgtt gtttttggtt aaacatcttt gtgaaaacat
1621 cctcctgtca tgtatttgct atattcaaat gttaaacccg tgcagaatag aacatataca
1681 aaaaaaaaca acacaacaca tttttaaaca ttattaaata tcaagtattg ctggcagttc
1741 tgtttctgtt ttacagtacc ctttgccaca gttctccgct tttcctggtc cagattccac
1801 aagtctgatt caccaatagc aaagcgaata aacaaccaaa gcagccaatc actgcttgta
1861 gactgtcctg cgagaccggc ccattccagc acattctgga aacttccttt atatgataat
1921 tataaataca tttaaattat tgatacaaaa catgtaattc ctagaacata accatagcaa
1981 tcattagttt tcagggtaat tatgtatttt taggatttga ctgcggaaag atctggtcat
2041 gtgacgtctc atgaacgtca cggccctggg tttctataaa tacagtagga ctctcgacca
2101 tcggcagatt tttcgaagaa gaagatcagt ttcaggagcc gtactgttcc gtttcaacgc
2161 aaatattaac ggtaagagcg aatttcctag tttgttttca tgccattctt taaaaccata
2221 gcgtattact ttaattatag taaactttcg ctttctttat tacaagagac gttttgtgtt
2281 gattctccgc ggacattttc ggtcagacaa tcagaaaatg accgcggagg accagtaact
2341 tgcattacac gtaagttaaa tcttcgtgta ttaaaatggt taggttgtta acgtcaaata
2401 ggttaccgtg tttgcgtgtg atcaggttgg ttttgttaga tttttgtcag tatttttaat
2461 ttatttgttt tagtttattt attttttttg ctgaatcata gtttgtgaac aaagaacccg
2521 gatgttacat acagtacagc cgccatgtta cagagagtta taacttaatc attttaaaaa
2581 taattttgcc ttacttttag tttgtcatgt tgagaaatga ggaaatgtta aaatgaggaa
2641 atatccaatt aatttaatat atcaaaataa tccatgatta caatgcactg aactggagaa
2701 aattaagatg ttttctagtg tcatgaaaca aatgtaagag atgtacattg tagatgtttt
2761 atgtcaagaa ttggctagtt gatgcagcat actggcgata ctcagttgta ataacagtaa
2821 cgttacatgt taatagacta ctgagtatgc tgttctgtct atgtatgctc tgtaagctac
2881 gagaaggact tttttaaaca gtaaagggtg caatattttt acaaattgaa ttaaataaag
2941 gctgtctatt aagtaatatg cttgatattt ttcttacttg atcggaaata agaaaaaata
3001 taaacgttgt tgctctaaaa atcctagttc agtttagcca accacaaata cctttttgtt
3061 cctccaacag tttttttttc ttctctataa tatttggcag tctatagtac tccaaatgtt
3121 tccccacagt ctaactaatt ggtacagcca aaatcatgac acttattgca ataataattt
3181 tggttcattg gcattgttga tagcctgtgc cactaatatg gtcgattgat catgcttcag
3241 gaagaaaact atattgtttg atgtaagatt attaaatctt cacctgcttc cattacaaac
3301 tattcccatc ttattgaatt ctggtatgtc ttaaaggatt agttcacttc ccaaatcaaa
3361 atttacttag ccttttttca tccatgatcc ctttttttca tcattaatga agaaattgtt
3421 tttgaaaaag tttcaagatt tttttctcta tattgtggag cttgttagtt taaaattcca
3481 aaatgcaata tgtggcttca aatggttcta aatgatccca gtcaaggaat aacagtctta
3541 tctaatgaaa ccattagacc tttaaaaata aaaaaaataa aagtatttat ttttaaatga
3601 ctgagtgatt aagttgaatt tcagcgtttc cttactgtgt agaagtcctt ccttactggc
3661 cccacccttt ggttctctgc caatctgcta cctaatgtaa tgttgtggaa cattattatt
3721 ctttatttct taatttttta ttttttattt taaaacaatg taaactgcac agatgtgcag
3781 tttgtttaaa atggccaatg ctttggaaat gcatgacata attagatttc atgatgcaca
3841 aagccaaatc tcagagcttg tgcaaaatga gctatcattt cactaggtaa gaccctaaat
3901 tttcatatag gatcatttgg acaattttgc tgcaggtaaa atgcattcta tagtccactg
3961 tcagccattg ttttggatag tatttatttt tctctacaag tatagtcaat agttttctat
4021 tattttaaag gtttgtaaca tttaagggtg accaaatgca aagtaaaatt tcattttcgg
4081 gtgaactatc tcgtttaaca tgggagaagt gcaaaacata cattattggc tagaacattg
4141 tagtattttt taaatggaaa tgtgtgattg ctaatcttac tttgaatttg tttacaggga
4201 tccaagctta taacttcgta tagcatacat tatacgaagt tatccggtcg ccaccatggt
4261 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
4321 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
4381 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
4441 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
4501 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
4561 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
4621 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
4681 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
4741 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
4801 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
4861 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
4921 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
4981 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
5041 aacctcccac acctccccgt gaacgtgaaa cataaaatga atgcaattgt tgttgttaac
5101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
5161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
5221 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
5281 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
5341 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
5401 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
5461 accctaatca agataacttc gtatagcata cattatacga agttataccg gtggatccaa
5521 aaagggcgaa ttcgacccaa gtttgtacaa aaaagttgaa cgagaaacgt aaaatgatat
5581 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
5641 acaacatatg cagtcactat gaaccaacta cttagatggt attagtgacc tgtagaatta
5701 attcgagctc tagagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg
5761 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
5821 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
5881 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
5941 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
6001 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
6061 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
6121 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
6181 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
6241 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
6301 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
6361 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
6421 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
6481 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
6541 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
6601 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
6661 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
6721 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
6781 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
6841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
6901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
6961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
7021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
7081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
7141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
7201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
7261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
7321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
7381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
7441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
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