| 质粒名称: | pT2-5xERE:GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 125690 |
| 存储实验室: | Daniel Gorelick, Marnie Halpern |
|
载体骨架基本信息 |
|
| 载体名称: | pTol2 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 斑马鱼表达 |
| 筛选标记: | -- |
| 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: | 5xERE:GFP |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 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: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pT2-5xERE:GFP was a gift from Daniel Gorelick & Marnie Halpern (Addgene plasmid # 125690 ; http://n2t.net/addgene:125690 ; RRID:Addgene_125690) |
| 参考文献部分: | Visualization of estrogen receptor transcriptional activation in zebrafish. Gorelick DA, Halpern ME. Endocrinology. 2011 Jul;152(7):2690-703. doi: 10.1210/en.2010-1257. Epub 2011 May 3. 10.1210/en.2010-1257 PubMed 21540282 |
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 pT2-5xERE:GFP 7205 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Transgene to report estrogen receptor activity in zebrafish.
ACCESSION .
VERSION .
KEYWORDS pT2-5xERE:GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7205)
AUTHORS Gorelick DA, Halpern ME
TITLE Visualization of estrogen receptor transcriptional activation in
zebrafish.
JOURNAL Endocrinology. 2011 Jul;152(7):2690-703. doi: 10.1210/en.2010-1257.
Epub 2011 May 3.
PUBMED 21540282
REFERENCE 2 (bases 1 to 7205)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7205
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(15..26)
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
regulatory 87..96
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 93..812
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 93..809
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(138..159)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(399..418)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 746..767
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 857..991
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(881..900)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 935..954
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1663..1679)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1663..1679)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(1716..1736)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1716..1734)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1755..1771)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1755..1771)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1768..1790)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1779..1795
/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(1803..1833)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1848..1869
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1986..2003)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2157..2745)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2237..2256)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2916..3776)
/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 3539..3558
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3777..3881)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3907..4362)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4044..4065)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4256..4275
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4489..4511
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4503..4520
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4504..4520
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4530..4549
/label=T7
/note="T7 promoter, forward primer"
promoter 4530..4548
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 cacccagctc tgctctgcag ctcccaccag tgtctacccc tggacccctt gccgggcttt
61 ccccaaactt cgaggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
121 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
181 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
241 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
301 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
361 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
421 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
481 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
541 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
601 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
661 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
721 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
781 tcactctcgg catggacgag ctgtacaagt aaagcggccg ccaccgcggt ggagctcgaa
841 ttaattcatc gatgatgatc cagacatgat aagatacatt gatgagtttg gacaaaccac
901 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
961 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
1021 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
1081 tatggctgat tatgatcctc tagatcagat ctgcgaagat acggccacgg gtgctcttga
1141 tcctgtggct gattttggac tgtgctgctc gcagctgctg atgaatcaca tacttcctcc
1201 attttcttcc actgattgac tgttataatt tccctaattt ccaggtcaag gtgctgtgca
1261 ttgtggtaat agatgtgaca tgacgtcact tccaaaggac caatgaacat gtctgaccaa
1321 tttcatataa tgtgaaaacg attttcatag gcagaataaa taacatttaa attaaactgg
1381 gcatcagcgc aattcaattg gtttggtaat agcaagggaa aatagaatga agtgatctcc
1441 aaaaaataag tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt
1501 ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag
1561 taaaaatccc caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt
1621 acacctctgg ttcttgaccc cctaccttca gcaagcccag cagatccact agttctagag
1681 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
1741 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
1801 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
1861 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
1921 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
1981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
2041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
2101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
2161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
2221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
2281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
2341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
2401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
2461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
2521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
2581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
2641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
2701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
2761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
2821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
2881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
2941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
3001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
3061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
3121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
3181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
3241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
3301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
3361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
3421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
3481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
3541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
3601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
3661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
3721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
3781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
3841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
3901 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
4381 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
4441 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
4501 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
4561 ccaaatagta ggaattaccc acctgtacaa gtgctgaaaa cttggatgaa taagcccgtt
4621 tgccattttc tactgctatt ttaaatcttt tctgtatttg tctgcatttg tctttaccct
4681 tgaataaatg ttttagttgt tttttttcaa ttatgactgt gtttaacaga cattattaca
4741 ataatatgta atagtagtac acactattat tatgtaatag tacttgttga ctgtatttga
4801 gaactggacc gagtgagtgt tacgtcaccc attcaaaatg acttacttct ggctccaaca
4861 aaatgaagtt aattcagttg ccatttttca ctgtatggac atcgccgtgt tggagctaga
4921 cgttgccaat aagcaagaaa gagccgagat gcgtcgagtc tgagtcaccg ttcctatggc
4981 aacccctcta accaatcaga agtaagcttg ttggaagtcc acagcctacc acttgaaagc
5041 gggctgcaca aaatctgtca aacgttttga acgttggatg tgagagcaca tacttttatt
5101 aaggcatctg gttggtcagt ttataatatc aacaacttgg gctacagaaa agaaaagtta
5161 ttacagaaat tatgattaac aagtacatgt taaataaaga ttttaatatg aatgccacca
5221 ctggagcatt catgccattt ggagcttctt cctgtttgga tcactagaag gaggaggtca
5281 ctcattacag ttctcatata cagtcgttgg ttggttggtt ggttggtaga ttgattgatt
5341 gattgattga ttgattgatt gattgattga ttgattgatt gattgattga ttgattgatt
5401 ggtagtcaaa ataagaaata atttccacag attcattaca gaaatgatta aatgcataca
5461 taaaaaactg gggggggggg ggatacaaca acacacttaa gtcacatttg cctacgtaaa
5521 gaaaagtaaa gaaaatcaat agctatattt tacatctcct ttttttgctg tcttttaatt
5581 agccttgttt tgctgttatc ttgattgaaa ctgtaacttc ttcacctgct tcttttcttt
5641 gtaggttttg ccagcccaga aacaggcatg gctgtgcaag gccagagcac catgcacaat
5701 gcgctgcatg tcttcatgaa cggctcaatg tcctcagtcc agggctcagc caacgacccc
5761 atcttcctcc ttcaccatgc tttcattgac aggtaacaaa cacgtcatga cattagactg
5821 cacagttttt gacaaagttc atacaatctg ttgtttatag ctgctacaat tagtgaagtt
5881 tgtgaatgta cttggatgag cagcgaaaga tcaattgaga tcaattgtta gagtttggtt
5941 gccctgcaga gcaaagaaca aaaaataatc tggtggcttt actgcgtgag gttattattg
6001 gtggaataga aacacaaaac ataattgcat ttatttgttt aattttttat cttatcttaa
6061 ctttcatctt gcatatttgt ttcttacatc atttctagca tctttgagcg ctggctaaga
6121 actcatcagc ctccccggtc catctaccca cgtaccaatg caccaattgg ccacaatgac
6181 ggctactaca tggtgccatt ccttcctctt tataggaatg gagactacct cctgtccaac
6241 aatgctcttg gatacgagta cgcctacctg ttggacccag gtcattgcac aacaccagaa
6301 atgccctctg atctgcaaaa gacgtgaata tctgttcaga cacccatatc cactctgttc
6361 cacacaggtc agaggtttgt ccaggagttc ttgacagagg tgtaaaaagt actcaaaaat
6421 tttactcaag tgaaagtaca agtacttagg gaaaatttta ctcaattaaa agtaaaagta
6481 tctggctaga atcttacttg agtaaaagta aaaaagtact ccattaaaat tgtacttgag
6541 tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga gattcttgtt taagctttta
6601 atctcaaaaa acattaaatg aaatgcatac aaggttttat cctgctttag aactgtttgt
6661 atttaattat caaactataa gacagacaat ctaatgccag tacacgctac tcaaagttgt
6721 aaaacctcag atttaacttc agtagaagct gattctcaaa attgttagtg tcaagcctag
6781 ctcttttggg gctgaaaagc aatcctgcag tgctgaaaag cctctcacag gcagccgatg
6841 cgggaagagg tgtattagtc ttgatagaga ggctgcaaat agcaggaaac gtgagcagag
6901 actccctggt gtctgaaaca caggccagat gggccctcga gtaggtcact gtgacctggt
6961 cgagtaggtc actgtgacct ggtcgagtag gtcactgtga cctggtcgag taggtcactg
7021 tgacctggtc gagtaggtca ctgtgacctg gtcgagatat ccagtgacgt aggaagtcca
7081 tccattcaca gcgcttctat aaaggcgcca gctgaggcgc ctactactcc aaccgcgact
7141 gcagcgagca actgagaaga ctggatagag ccggcggttc cgcgaacgag cagtgaccgc
7201 gctcc
//
