| 质粒名称: | pNP1 OR sfGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107364 |
| 存储实验室: | James Collins |
|
载体骨架基本信息 |
|
| 载体名称: | pNP1 |
| 载体出品公司: | New England Biolabs |
| 空载体大小: | 2202 bp |
| 完整质粒大小: | 3049 bp |
| 载体类型: | 细菌细胞表达 |
| 筛选标记: | 新霉素(G418) |
| 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: | OR gate + sfGFP |
| 插入大小: | 877 bp |
| 启动子: | T7 |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | CCCGCGAAATTAATACGACTCACTATAGG |
| 3’测序引物 | AGTGCCAAGCTTCCGGATATAGTTC |
|
文献引用方法 |
|
| 材料和方法部分: |
pNP1 OR sfGFP was a gift from James Collins (Addgene plasmid # 107364 ; http://n2t.net/addgene:107364 ; RRID:Addgene_107364)
|
| 参考文献部分: | BioBits Explorer: A modular synthetic biology education kit. Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ. Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. eCollection 2018 Aug. 10.1126/sciadv.aat5105 PubMed 30083608 |
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pNP1 OR sfGFP载体质粒基本信息
pNP1 OR sfGFP质粒图谱载体图谱和pNP1 OR sfGFP载体序列质粒序列多克隆位点信息

pNP1 OR sfGFP质粒载体简介
pNP1 OR sfGFP质粒序列载体序列
LOCUS pNP1 OR sfGFP 3079 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses sfGFP (a GFP) in Ecoli off a T7 promoter when pNP1 OR
input A and pNP1 OR input B are co-expressed.
ACCESSION .
VERSION .
KEYWORDS pNP1 OR sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3079)
AUTHORS Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T,
Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
TITLE BioBits Explorer: A modular synthetic biology education kit.
JOURNAL Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105.
eCollection 2018 Aug.
PUBMED 30083608
REFERENCE 2 (bases 1 to 3079)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3079
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 26..45
/label=T7
/note="T7 promoter, forward primer"
promoter 26..44
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 847..894
/codon_start=1
/product="11th beta-strand of superfolder GFP"
/label=GFP11
/note="associates with GFP(1-10), consisting of the first
10 beta-strands, to reconstitute fluorescent GFP (Cabantous
et al., 2005; Kamiyama et al., 2016)"
/translation="RDHMVLHEYVNAAGIT"
CDS 895..918
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
primer_bind complement(1057..1075)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1071..1118
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin complement(1336..1924)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1416..1435)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2096..2956)
/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 2719..2738
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2957..3061)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggatgaa cttaagatcg
61 cgctttatgg ttatgaaaca gaggagacat aacatgaaag cgcgaaacag aacaagaata
121 agtgcaaatg cttgttctat cgtcttgaaa cagaggagac aagacatgag aacaagcaac
181 ctggcggcag cgcaaaagat gagcaaaggt gaagaactgt ttaccggcgt tgtgccgatt
241 ctggtggaac tggatggcga tgtgaacggt cacaaattca gcgtgcgtgg tgaaggtgaa
301 ggcgatgcca cgattggcaa actgacgctg aaatttatct gcaccaccgg caaactgccg
361 gtgccgtggc cgacgctggt gaccaccctg acctatggcg ttcagtgttt tagtcgctat
421 ccggatcaca tgaaacgtca cgatttcttt aaatctgcaa tgccggaagg ctatgtgcag
481 gaacgtacga ttagctttaa agatgatggc aaatataaaa cgcgcgccgt tgtgaaattt
541 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca cggattttaa agaagatggc
601 aatatcctgg gccataaact ggaatacaac tttaatagcc ataatgttta tattacggcg
661 gataaacaga aaaatggcat caaagcgaat tttaccgttc gccataacgt tgaagatggc
721 agtgtgcagc tggcagatca ttatcagcag aataccccga ttggtgatgg tccggtgctg
781 ctgccggata atcattatct gagcacgcag accgttctgt ctaaagatcc gaacgaaaaa
841 ggcacgcggg accacatggt tctgcacgaa tatgtgaatg cggcaggtat tacgtggagc
901 catccgcagt tcgaaaaata aaggatcccg ggaattctcg agtaaggtta acctgcagga
961 ggcctttaat taaggtggtg cggccgcgct agcggtcccg ggggatcgat ccggctgcta
1021 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
1081 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
1141 gaagcttggc actggccgac cggggtcgag cactgactcg ctgcgctcgg tcgttcggct
1201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
1261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
1321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
1381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
1441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
1501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
1561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
1621 cgccttatcc ggtaactatc gtcttgagtc caacccgcta agacacgact tatcgccact
1681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
1741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
1801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
1861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
1921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcaca
1981 gatccgggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
2041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
2101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
2161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
2221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
2281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
2341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
2401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
2461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
2521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
2581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
2641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
2701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
2761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
2821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
2881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
2941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
3001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
3061 gcacatttcc ccgaaaagt
//
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pNP1 OR sfGFP质粒载体应用举例
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