| 质粒名称: | 35S-eGFP-nosT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80127 |
| 存储实验室: | Yutaka Kodama |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18 |
| 载体出品公司: | Clontech |
| 空载体大小: | 2686bp |
| 完整质粒大小: | 4144bp |
| 载体类型: | Plant Expression |
| 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 |
|
| 启动子: | CaMV35S |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | GGATCCATGGTGAGCAAGGGCGAGGAGCTG |
| 3’测序引物 | TTACTTGTACAGCTCGTCCATGCCGAGAGT |
|
文献引用方法 |
|
| 材料和方法部分: | 35S-eGFP-nosT was a gift from Yutaka Kodama (Addgene plasmid # 80127 ; http://n2t.net/addgene:80127 ; RRID:Addgene_80127) |
| 参考文献部分: | In planta comparative analysis of improved green fluorescent proteins with reference to fluorescence intensity and bimolecular fluorescence complementation ability. Fujii Y, Kodama Y. Plant Biotech. 2015; 32(1): 81-87. 10.5511/plantbiotechnology.15.0120a |
买家导航
35S-eGFP-nosT载体质粒基本信息
35S-eGFP-nosT质粒图谱载体图谱和35S-eGFP-nosT载体序列质粒序列多克隆位点信息
35S-eGFP-nosT质粒载体简介
35S-eGFP-nosT质粒序列载体序列
LOCUS 35S-eGFP-nosT 4144 bp DNA circular SYN 14-APR-2025
DEFINITION Transient expression vector for eGFP in plants.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4144)
AUTHORS Fujii Y, Kodama Y
TITLE In planta comparative analysis of improved green fluorescent
proteins with reference to fluorescence intensity and bimolecular
fluorescence complementation ability.
JOURNAL Plant Biotech. 2015; 32(1): 81-87.
REFERENCE 2 (bases 1 to 4144)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Biotech."; date: "2015"; volume: " 32(1)"; pages: " 81-87"
COMMENT Sequence Label: 35S-eGFP-nosT
FEATURES Location/Qualifiers
source 1..4144
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 31..48
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 165..186
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(201..206,207..224,225..231)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 239..255
/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)."
primer_bind 244..266
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 263..279
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 263..279
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
terminator complement(317..569)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(join(588..1301,1302..1304,1305..1307))
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
CDS complement(591..1307)
/codon_start=1
/product="green fluorescent protein"
/label=GFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(633..654)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 982..1001
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1241..1262
/label=EGFP-N
/note="EGFP, reverse primer"
misc_feature 1327..1382
/label=TMV Omega
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
promoter complement(1402..1747)
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind complement(1439..1459)
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
primer_bind complement(1807..1824)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1807..1823)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1816..1838)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2032..2051)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2151..2173
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2211..2229)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2297..2401
/gene="bla"
/label=AmpR promoter
CDS join(2402..2470,2471..3262)
/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 complement(2620..2639)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3433..4021
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3922..3941
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
61 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
241 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttctcatgtt
301 tgacagctta tcatcggatc tagtaacata gatgacaccg cgcgcgataa tttatcctag
361 tttgcgcgct atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc
421 ataaaaaccc atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg
481 taattcaaca gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag
541 aaactttatt gccaaatgtt tgaacgatct gcagccgggc ggccgcttta cttgtacagc
601 tcgtccatgc cgtgagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
661 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc
721 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
781 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
841 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
901 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
961 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
1021 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
1081 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
1141 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
1201 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
1261 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatgga tcctctagag
1321 tcgactgtaa ttgtaaatag taattgtaat gttgtttgtt gtttgttgtt gttggtaatt
1381 gttgaaggat ctcaagaagc ctgtcctctc caaatgaaat gaacttcctt atatagagga
1441 agggtcttgc gaaggatagt gggattgtgc gtcatccctt acgtcagtgg agatatcaca
1501 tcaatccact tgctttgaag acgtggttgg aacgtcttct ttttccacga tgttcctcgt
1561 gggtgggggt ccatctttgg gaccactgtc ggtagaggca tcttgaacga tagcctttcc
1621 tttatcgcaa tgatggcatt tgtaggagcc accttccttt tctactgtcc tttcgatgaa
1681 gtgacagata gctgggcaat ggaatccgag gaggtttccc gatattaccc tttgttgaaa
1741 agtctcaata gccctctggt cttctgaggg gggatcctct agagcctgca ggcatgcaag
1801 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
1861 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
1921 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
1981 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
2041 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
2101 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
2161 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
2221 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
2281 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
2341 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
2401 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
2461 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
2521 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
2581 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
2641 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
2701 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
2761 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
2821 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
2881 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
2941 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
3001 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
3061 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
3121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
3181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
3241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
3301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
3361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
3421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
3481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
3541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
3601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
3661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
3721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
3781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
3841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
3901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
3961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
4021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
4081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
4141 tgat
//
