| 质粒名称: | pFE-sfGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 132998 |
| 存储实验室: | David Savage |
|
载体骨架基本信息 |
|
| 载体名称: | pFE |
| 空载体大小: | 3132bp |
| 完整质粒大小: | 3871 bp |
| 载体类型: | 细菌细胞表达 |
| 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 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | Synthetic |
| 插入大小: | 714 bp |
| 标签/融合蛋白: | 6xhis (C terminal on insert) |
| 克隆方法: | 未知 |
| 5’测序引物 | CCTTTCGTCTTCACCTCGAG |
|
文献引用方法 |
|
| 材料和方法部分: |
pFE-sfGFP was a gift from David Savage (Addgene plasmid # 132998 ; http://n2t.net/addgene:132998 ; RRID:Addgene_132998)
|
| 参考文献部分: | DABs are inorganic carbon pumps found throughout prokaryotic phyla. Desmarais JJ, Flamholz AI, Blikstad C, Dugan EJ, Laughlin TG, Oltrogge LM, Chen AW, Wetmore K, Diamond S, Wang JY, Savage DF. Nat Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi: 10.1038/s41564-019-0520-8. 10.1038/s41564-019-0520-8 PubMed 31406332 |
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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 pFE-sfGFP 3871 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION pFE carrying a sfGFP gene.
ACCESSION .
VERSION .
KEYWORDS pFE-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3871)
AUTHORS Desmarais JJ, Flamholz AI, Blikstad C, Dugan EJ, Laughlin TG,
Oltrogge LM, Chen AW, Wetmore K, Diamond S, Wang JY, Savage DF
TITLE DABs are inorganic carbon pumps found throughout prokaryotic phyla.
JOURNAL Nat Microbiol. 2019 Aug 12. pii: 10.1038/s41564-019-0520-8. doi:
10.1038/s41564-019-0520-8.
PUBMED 31406332
REFERENCE 2 (bases 1 to 3871)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3871
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(236..859)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 767..786
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter complement(860..964)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1140..1728)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1220..1239)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 1816..1910
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1941..2735)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2052..2071)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2662..2681
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2914..2932
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2953..3026
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 2953..2971
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2978..2996
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 3036..3047
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3054..3773
/codon_start=1
/product="enhanced GFP (EGFP) derivative that exhibits
robust folding and enhanced photostability (Zhong et al.,
2019)"
/label=SiriusGFP
/note="mammalian codon-optimized"
/translation="MSVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLT
LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKD
DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGI
KANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVL
LEFVTAAGITLGMDELYK"
primer_bind complement(3102..3123)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 3710..3731
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3780..3797
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 3848..63
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
61 aatccgccgc cgctagcagt tcgtagacgg aaacaaacgc agaatccaag caaaagggaa
121 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
181 tttatcaggg ttatgtccac tgggttcgtg ccttcatccg gatatccagg aactgttaag
241 acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca aggccgaata
301 agaaggctgg ctctgcacct tggtgatcaa ataattcgat agcttgtcgt aataatggcg
361 gcatactatc agtagtaggt gtttcccttt cttctttagc gacttgatgc tcttgatctt
421 ccaatacgca acctaaagta aaatgcccca cagcgctgag tgcatataat gcattctcta
481 gtgaaaaacc ttgttggcat aaaaaggcta attgattttc gagagtttca tactgttttt
541 ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg atgacttagt aaagcacatc
601 taaaactttt agcgttatta cgtaaaaaat cttgccagct ttccccttct aaagggcaaa
661 agtgagtatg gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa gcccgcttat
721 tttttacatg ccaatacaat gtaggctgct ctacacctag cttctgggcg agtttacggg
781 ttgttaaacc ttcgattccg acctcattaa gcagctctaa tgcgctgtta atcactttac
841 ttttatctaa tctagacata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
961 cgcgtttccc tcgacaattc gcgctagacc tagggcgttc ggctgcggcg agcggtatca
1021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
1081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
1141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
1201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
1261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
1321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
1381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
1441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
1501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
1561 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
1621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
1681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
1741 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
1801 atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
1861 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
1921 ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
1981 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
2041 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
2101 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
2161 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
2221 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
2281 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
2341 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
2401 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
2461 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
2521 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
2581 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
2641 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
2701 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
2761 tgatcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt
2821 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtctaa
2881 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
2941 cttcacctcg agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact
3001 gagcacatca gcaggacgca ctgaccgaat tcattaaaga ggagaaaggt accatgtccg
3061 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
3121 acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc accaacggca
3181 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
3241 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
3301 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atctccttca
3361 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
3421 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
3481 tggagtacaa cttcaacagc cacaacgtct atatcacggc cgacaagcag aagaacggca
3541 tcaaggcgaa cttcaagatc cgccacaacg tcgaggacgg cagcgtgcag ctcgccgacc
3601 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
3661 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
3721 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagagcggtc
3781 accatcacca tcaccattaa gaattcctgc agcccggggg atcccatggt acgcgtgcta
3841 gaggcatcaa ataaaacgaa aggctcagtc g
//
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