| 质粒名称: | pFPV25.1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20668 |
| 存储实验室: | Raphael Valdivia |
|
载体骨架基本信息 |
|
| 载体名称: | pFPV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4764 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
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 | -- |
| 标签/融合蛋白: | GFP mut3（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI (未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pFPV25.1 was a gift from Raphael Valdivia (Addgene plasmid # 20668 ; http://n2t.net/addgene:20668 ; RRID:Addgene_20668) |
| 参考文献部分: | Bacterial genetics by flow cytometry: rapid isolation of Salmonella typhimurium acid-inducible promoters by differential fluorescence induction. Valdivia RH, Falkow S. Mol Microbiol. 1996 Oct . 22(2):367-78. 10.1046/j.1365-2958.1996.00120.x PubMed 8930920 |
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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 pFPV25.1 5395 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION S. typhimurium rpsM promoter driving gfpmut3 expression in bacteria.
Promoter is not regulated by acidic conditions. .
ACCESSION .
VERSION .
KEYWORDS pFPV25.1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5395)
AUTHORS Valdivia RH, Falkow S
TITLE Bacterial genetics by flow cytometry: rapid isolation of Salmonella
typhimurium acid-inducible promoters by differential fluorescence
induction.
JOURNAL Mol Microbiol. 1996 Oct . 22(2):367-78.
PUBMED 8930920
REFERENCE 2 (bases 1 to 5395)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5395
/organism="synthetic DNA construct"
/mol_type="other DNA"
oriT 1303..1390
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS complement(2020..2736)
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(2063..2083)
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind 2678..2706
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind complement(3404..3422)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3490..3594
/gene="bla"
/label=AmpR promoter
CDS 3595..4455
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3813..3832)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4626..5214
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5115..5134
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5368..5385
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgctcaca gcccatggcc ttgagctgtc ggcctatggc ctgcaaagtc ctgtcgttct
61 tcatcgggcc accaagcgca gccagatcga gccgtcctcg gttgtcagtg gcgtcaggtc
121 gagcaagagc aacgatgcga tcagcagcac caccgtaggc atcatggaag ccagcatcac
181 ggttagccat agcttccagt gccacccccg cgacgcgctc cgggcgctct gcgcggcgct
241 gctcacctcg gcggctacct cccgcaactc tttggccagc tccacccatg ccgcccctgt
301 ctggcgctgg gctttcagcc actccgccgc ctgcgcctcg ctggcctgct gggtctggct
361 catgacctgc cgggcttcgt cggccagtgt cgccatgctc tgggccagcg gttcgatctg
421 ctccgctaac tcgttgatgc ctctggattt cttcactctg tcgattgcgt tcatggtcta
481 ttgcctcccg gtattcctgt aagtcgatga tctgggcgtt ggcggtgtcg atgttcaggg
541 ccacgtctgc ccggtcggtg cggatgcccc ggccttccat ctccaccacg ttcggcccca
601 ggtgaacacc gggcaggcgc tcgatgccct gcgcctcaag tgttctgtgg tcaatgcggg
661 cgtcgtggcc agcccgctct aatgcccggt tggcatggtc ggcccatgcc tcgcgggtct
721 gctcaagcca tgccttgggc ttgagcgctt cggtcttctg tgccccgccc ttctccgggg
781 tcttgccgtt gtaccgcttg aaccactgag cggcgggccg ctcgatgccg tcattgatcc
841 gctcggagat catcaggtgg cagtgcgggt tctcgccgcc accggcatgg atggccagcg
901 tatacggcag gcgctcggca ccggtcaggt gctgggcgaa ctcggacgcc agcgccttct
961 gctggtcgag ggtcagctcg accggcaggg caaattcgac ctccttgaac agccgcccat
1021 tggcgcgttc atacaggtcg gcagcatccc agtagtcggc gggccgctcg acgaactccg
1081 gcatgtgccc ggattcggcg tgcaagactt catccatgtc gcgggcatac ttgccttcgc
1141 gctggatgta gtcggccttg gccctggccg attggccgcc cgacctgctg ccggttttcg
1201 ccgtaaggtg ataaatcgcc atgctgcctc gctgttgctt ttgcttttcg gctccatgca
1261 atggccctcg gagagcgcac cgcccgaagg gtggccgtta ggccagtttc tcgaagagaa
1321 accggtaagt gccccctccc ctacaaagta gggtcgggat tgccgccgct gtgcctccat
1381 gatagcctac gagacagcac attaacaatg gggtgtcaag atggttaagg ggagcaacaa
1441 ggcggcggat cggctggcca agctcgaaga acaacgagcg cgaatcaatg ccgaaattca
1501 gcgggtgcgg gcaagggaac agcagcaaga gcgcaagaac gaaacaaggc gcaaggtgct
1561 ggtgggggcc atgattttgg ccaaggtgaa cagcagcgag tggccggagg atcggctcat
1621 ggcggcaatg gatgcgtacc ttgaacgcga ccacgaccgc gccttgttcg gtctgccgcc
1681 acgccagaag gatgagccgg gctgaatgat cgaccgagac aggccctgcg gggctgcaca
1741 cgcgccccca cccttcgggt agggggaaag gccgctaaag cggctaaaag cgctagcagc
1801 acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt
1861 accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg
1921 agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
1981 ctaccgcatt aaagcttgca tgcctgcagg tctggacatt tatttgtata gttcatccat
2041 gccatgtgta atcccagcag ctgttacaaa ctcaagaagg accatgtggt ctctcttttc
2101 gttgggatct ttcgaaaggg cagattgtgt ggacaggtaa tggttgtctg gtaaaaggac
2161 agggccatcg ccaattggag tattttgttg ataatggtct gctagttgaa cgcttccatc
2221 ttcaatgttg tgtctaattt tgaagttaac tttgattcca ttcttttgtt tgtctgccat
2281 gatgtataca ttgtgtgagt tatagttgta ttccaatttg tgtccaagaa tgtttccatc
2341 ttctttaaaa tcaatacctt ttaactcgat tctattaaca agggtatcac cttcaaactt
2401 gacttcagca cgtgtcttgt agttcccgtc atctttgaaa aatatagttc tttcctgtac
2461 ataaccttcg ggcatggcac tcttgaaaaa gtcatgctgt ttcatatgat ctgggtatct
2521 cgcaaagcat tgaacaccat aaccgaaagt agtgacaagt gttggccatg gaacaggtag
2581 ttttccagta gtgcaaataa atttaagggt aagttttccg tatgttgcat caccttcacc
2641 ctctccactg acagaaaatt tgtgcccatt aacatcacca tctaattcaa caagaattgg
2701 gacaactcca gtgaaaagtt cttctccttt actcatatgt atatctcctt cttaaatcta
2761 gaggatctta acattttcag cgatacccgc tgcagccagg atggctttag aacgggtctt
2821 gccgacaccg tagatcgaag ttaacgcgat cacggcgtgt ttctgatcag gaatgttaat
2881 gcctgctata cgggccacta tgcactccta ctatttaata tgtacgtacc atgctgaaaa
2941 gcccgttttc aggatactca aatggaaacg tacagacata caaaagattg gctggctaat
3001 ctagccagct caacccaact ttgcaagaaa aatatgcgaa aaaatcagcc ttggcgctgt
3061 ttatgcttcg gctcggcact gcaaatcaca cggatgacac catcacgctt aacgattttg
3121 cagttacggc ataatttctt gacggaagca cgaactttca tttttactct ccgtaacttc
3181 tcaggcgacc aattaacggc cgtagccttt caggttcgcc ttcttcaatg cagactcata
3241 ctgactggac atcatcagag tttgcacttg agccataaag tccataatca cgacaacaac
3301 gataagcagt gaggtcccac cgaagtagaa cggcactttc attgcatcac gcatgaactc
3361 cgggatcccc gggtaccgag ctcgaattct tgaagacgaa agggcctcgt gatacgccta
3421 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
3481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
3541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
3601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
3661 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
3721 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
3781 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
3841 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
3901 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
3961 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
4021 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
4081 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
4141 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
4201 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
4261 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
4321 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
4381 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
4441 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
4501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
4561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
4621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
4681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
4741 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
4801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
4861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
4921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
4981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
5041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
5101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
5161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
5221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
5281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
5341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
