| 质粒名称: | 35S-sfGFP-nosT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80129 |
| 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: | 2686 bp |
| 完整质粒大小: | 4180 bp |
| 载体修饰: | -- |
| 载体类型: | 植物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CaMV35S |
| 标签/融合蛋白: | sfGFP |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 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-sfGFP-nosT was a gift from Yutaka Kodama (Addgene plasmid # 80129 ; http://n2t.net/addgene:80129 ; RRID:Addgene_80129) |
| 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
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 35S-sfGFP-nosT 4180 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Transient expression vector for sfGFP in plants.
ACCESSION .
VERSION .
KEYWORDS 35S-sfGFP-nosT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4180)
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 4180)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4180
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 21..365
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 308..328
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
misc_feature 385..440
/label=TMV Omega
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
CDS 460..1179
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(505..526)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1113..1134
/label=EGFP-C
/note="EGFP, forward primer"
terminator 1198..1450
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(1488..1504)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1523..1539)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1536..1558)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1547..1563
/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(1571..1601)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1616..1637
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1754..1771)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1925..2513)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2005..2024)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2684..3544)
/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 3307..3326
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3545..3649)
/gene="bla"
/label=AmpR promoter
primer_bind 3717..3735
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3773..3795)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3895..3914
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4108..4130
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4122..4139
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4123..4139
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 ctcagaagac cgagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc
61 ggattccatt gcccagctat ctgtcactta tcgaaaggac agtagaaaag gaaggtggct
121 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctaccgaca
181 gtggtcccaa agatggaccc ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa
241 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
301 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
361 ggacaggctt cttgagatcc ttcaacaatt accaacaaca acaaacaaca aacaacatta
421 caattactat ttacaattac agtcgactct agaggatcca tggtgagcaa gggcgaggag
481 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
541 ttcagcgtga gaggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
601 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
661 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
721 gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga cggcacttac
781 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
841 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
901 agccacaacg tctatatcac tgccgacaag cagaagaacg gcatcaaggc caacttcaag
961 atccgccaca acgttgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
1021 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtt
1081 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
1141 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgccc ggctgcagat
1201 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
1261 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
1321 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
1381 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
1441 ttactagatc cgatgataag ctgtcaaaca tagaattcgt aatcatggtc atagctgttt
1501 cctgtgtgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
2101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
2341 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
2701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
2761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
2821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
2881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
2941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
3001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
3061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
3121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
3181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
3241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
3301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
3361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
3421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
3481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
3541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
3601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
3661 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
3721 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
3781 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
3841 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
3901 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
3961 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
4021 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
4081 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
4141 ccaagcttgc atgcctgcag gtcgatctag aggatccccc
//
