| 质粒名称: | pSCM001 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 124605 |
| 存储实验室: | David Summers |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD TOPO |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 4127 bp |
| 完整质粒大小: | 5433bp |
| 载体类型: | 细菌表达 |
| 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: | mCherry-pHluorin、Cherry-Super Ecliptic pHluorin |
| 物种来源: | E. coli |
| 插入大小: | 1440 bp |
| 启动子: | pDAB |
| 标签/融合蛋白: | mCherry-(Gly-Gly-Ser)x2-pHluorin translational fusion protein |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | ACATGGCATGGATGAACTATACAAATGAAAACGGTCTCCAGCTTGGCT |
| 3’测序引物 | TCGCCATGTTGTCTTCTTCGCCTTTTGAAACCATGGTATGTATATCTCCTTCTTAAAGTTAAACA |
|
文献引用方法 |
|
| 材料和方法部分: | pSCM001 was a gift from David Summers (Addgene plasmid # 124605 ; http://n2t.net/addgene:124605 ; RRID:Addgene_124605) |
| 参考文献部分: | Indole Pulse Signalling Regulates the Cytoplasmic pH of E. coli in a Memory-Like Manner. Zarkan A, Cano-Muniz S, Zhu J, Al Nahas K, Cama J, Keyser UF, Summers DK. Sci Rep. 2019 Mar 7;9(1):3868. doi: 10.1038/s41598-019-40560-3. 10.1038/s41598-019-40560-3 PubMed 30846797 |
买家导航
pSCM001载体质粒基本信息
pSCM001质粒图谱载体图谱和pSCM001载体序列质粒序列多克隆位点信息
pSCM001质粒载体简介
pSCM001质粒序列载体序列
LOCUS sequence-242761- 5432 bp DNA circular SYN 15-DEC-2025
DEFINITION Expresses mCherry-pHluorin fusion protein in E. coli, a fluorescent
reporter for cytoplasmic pH.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5432)
AUTHORS Zarkan A, Cano-Muniz S, Zhu J, Al Nahas K, Cama J, Keyser UF,
Summers DK
TITLE Indole Pulse Signalling Regulates the Cytoplasmic pH of E. coli in a
Memory-Like Manner.
JOURNAL Sci Rep. 2019 Mar 7;9(1):3868. doi: 10.1038/s41598-019-40560-3.
PUBMED 30846797
REFERENCE 2 (bases 1 to 5432)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep.";
date: "2019-03-7"; volume: "9(1):3868. doi"; pages: "
10.1038/s41598-019-40560-3"
COMMENT Sequence Label: pSCM001
FEATURES Location/Qualifiers
source 1..5432
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(49..66)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 219..305
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 397..424
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 443..534
/gene="bla"
/label=AmpR promoter
CDS join(535..603,604..1395)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(753..772)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1566..2154
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2055..2074
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2308..2325
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2340..2480
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2406..2425)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2566..2588
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2744..3622)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPTDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 3649..3933
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 3856..3875
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 3960..3982
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(4138..4156)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 4406..4425
/label=mCherry-F
/note="mCherry, forward primer"
CDS 4719..5432
/codon_start=1
/product="pH-sensitive mutant of green fluorescent protein
(Ng et al., 2002)"
/label=superecliptic pHluorin
/note="excited strongly by blue light at neutral pH but
minimally fluorescent at acidic pH"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(4746..4774)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 5369..5389
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
1 aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
61 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
121 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
181 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
241 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
301 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
361 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
421 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
481 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
541 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
601 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
661 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
721 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
781 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
841 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
901 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
961 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
1021 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1081 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1141 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1201 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1261 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1321 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1381 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1441 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
1501 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
1561 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
1621 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
1681 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
1741 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
1801 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
1861 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
1921 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
1981 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2041 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2101 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
2161 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2221 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2281 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2341 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
2401 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
2461 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
2521 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
2581 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
2641 aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
2701 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
2761 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
2821 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
2881 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct
2941 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
3001 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
3061 cgctgatgta ctgacaagcc tcgcgtaccc gattatccgt cggtggatgg agcgactcgt
3121 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
3181 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
3241 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
3301 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
3361 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
3421 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
3481 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
3541 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
3601 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt
3661 gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca
3721 aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa
3781 aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg
3841 gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac
3901 gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct agaaataatt
3961 ttgtttaact ttaagaagga gatatacata ccatggtttc aaaaggcgaa gaagacaaca
4021 tggcgattat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
4081 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
4141 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
4201 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
4261 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
4321 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
4381 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
4441 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
4501 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
4561 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
4621 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tctaccggcg
4681 gtatggatga actgtacggc ggctctggcg gctctataag taaaggagaa gaacttttca
4741 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
4801 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
4861 ctactggaaa actacctgtt ccttggccaa cacttgtcac tactttaact tatggtgttc
4921 aatgcttttc aagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc
4981 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
5041 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
5101 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aacgatcacc
5161 aggtgtacat catggcagac aaacaaaaga atggaatcaa agctaacttc aaaattagac
5221 acaacattga agatggaggc gttcaactag cagaccatta tcaacaaaat actccaattg
5281 gcgatgggcc cgtcctttta ccagacaacc attacctgtt tacaacttct actctttcga
5341 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
5401 ttacacatgg catggatgaa ctatacaaat ga
//
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