| 质粒名称: | pBLADE(FP6**)-mCherry | |
|---|---|---|
| 出品公司: | Addgene | |
| 目录编号: | 168049 | |
| 存储实验室: | Barbara Di Ventura | |
|
载体骨架基本信息 |
||
| 载体名称: | pBAD33 | |
| 载体出品公司: | Clontech | |
| 空载体大小: | 4038 bp | |
| 完整质粒大小: | 4989 bp | |
| 载体类型: | 哺乳细胞表达 | |
| 载体抗性: | Chloramphenicol, 25 μg/mL | |
| 生长温度: | 37 ℃ | |
| 宿主菌: | DH5a | |
| 拷贝数: | 低拷贝 | |
|
插入基因信息 |
||
| 插入基因名称:11 | Blue light-inducible AraC dimers in Escherichia coli、BLADE | |
| 物种来源: | Synthetic | |
| 插入大小: | 828 bp | |
| 启动子: | J23101** | |
| 克隆方法: | Gibson Cloning | |
| 5’测序引物: | TCCCCATCGGTGATGTC | |
|
mCherry | |
| 物种来源: | Discosoma sp. | |
| 插入大小: | 708bp | |
| 启动子: | PBAD | |
| 克隆方法: | Restriction Enzyme | |
| 3’克隆位点: | SacI (not destroyed) | |
| 克隆方法: | HindIII (destroyed during cloning) | |
| 5’测序引物 | ATGCCATAGCATTTTTATCC | |
| 3’测序引物 | CTGATTTAATCTGTATCAGG | |
|
文献引用方法 |
||
| 材料和方法部分: | ||
| 参考文献部分: | ||
买家导航
pBLADE(FP6**)-mCherry载体质粒基本信息
pBLADE(FP6**)-mCherry质粒图谱载体图谱和pBLADE(FP6**)-mCherry载体序列质粒序列多克隆位点信息
pBLADE(FP6**)-mCherry质粒载体简介
pBLADE(FP6**)-mCherry质粒序列载体序列
LOCUS pBLADE(FP6**)-mCherry 6128 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses the synthetic gene BLADE FP6 under the constitutive
J23101** promoter. mCherry expression is controlled by the PBAD
promoter..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6128)
AUTHORS Romano E, Baumschlager A, Akmeric EB, Palanisamy N, Houmani M,
Schmidt G, Ozturk MA, Ernst L, Khammash M, Di Ventura B
TITLE Engineering AraC to make it responsive to light instead of
arabinose.
JOURNAL Nat Chem Biol. 2021 Apr 26. pii: 10.1038/s41589-021-00787-6. doi:
10.1038/s41589-021-00787-6.
PUBMED 33903769
REFERENCE 2 (bases 1 to 6128)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem
Biol. 2021 Apr 26. pii: 10.1038/s41589-021-00787-6. doi:
10.1038/s41589-021-00787-6."
COMMENT Sequence Label: pBLADE(FP6**)-mCherry
FEATURES Location/Qualifiers
source 1..6128
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(713..1372)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1287..1306
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1373..1475)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(1867..1884)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2001..2546)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(2081..2100)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2622..2641
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 3799..4083
/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 4006..4025
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 4138..4848
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(4283..4301)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 4551..4570
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(4887..4904)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 5057..5143
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5235..5262
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 5281..5372
/gene="bla"
/label=AmpR promoter
rep_origin join(5830..6128,1..157)
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5917..5936)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind join(6127..6128,1..20)
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
61 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
121 aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct
181 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
241 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
301 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
361 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
421 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
481 tctgctaatc ctgttaccag tcaggcattt gagaagcaca cggtcacact gcttccggta
541 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
601 ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
661 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
721 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
781 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
841 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
901 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
961 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
1021 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
1081 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
1141 tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
1201 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
1261 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
1321 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
1381 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
1441 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
1501 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
1561 ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt
1621 agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc
1681 tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag
1741 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
1801 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
1861 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
1921 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
1981 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
2041 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
2101 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
2161 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
2221 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
2281 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
2341 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
2401 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
2461 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
2521 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
2581 aggctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
2641 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
2701 atctgctcat gtttgacagc ttatcatttt tttaggctac cgatgtagcc taacgcgatc
2761 gatttatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
2821 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
2881 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
2941 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
3001 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
3061 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
3121 cggtggatgg agcgactcgt tttccgtttc gcactggaaa cccatgctgt accggtattc
3181 ccctgtttca tctcgcaccg gaatcatcgt caagaagttg acaaaccgtt ggccgttctt
3241 cttaaaattg accacctcaa cctgcacctc ggcgttccta tcaatcgctt tcctcatcgt
3301 attgatcgtg ttggagtcga cgtacttcct tgtcgatttc ggcttgacca ttccgtcggg
3361 tgactgaaga aaacggcagt ttctccccaa gacctccgca ttgctgtatc ctgtcatata
3421 gagaaaagct tccgaggcgt acacaattgg cgtgtctttt tgcttcaggt cgcacagaat
3481 cagagcaact gacgtgtcaa caggtcccag ttctacttgg gggtttggcc tcttcataat
3541 ctgaatcaga tagcccataa tgtcataacc gccgggagcg tagagcgtat gcatggtata
3601 tctccttctc tagtctctag tagctagcat atacctagga ctgagctagt gtaaactcta
3661 gaaatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
3721 ccgtcaagcc gtcaattgtc tgattcgtta ccaacggatg cgccatgaca ggacttttca
3781 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
3841 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
3901 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
3961 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
4021 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
4081 catacccgtt tttttgggct agcgaattcg agctcggtac ctaaggagga tttaaagatg
4141 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
4201 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
4261 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
4321 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
4381 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
4441 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctcctt gcaggacggc
4501 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
4561 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
4621 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
4681 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
4741 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
4801 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaaa gcttggctgt
4861 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
4921 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
4981 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
5041 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
5101 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
5161 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
5221 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
5281 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
5341 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
5401 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
5461 aagtaaaaga tgctgaagat cagttgggtg cagcaaacta ttaactggcg aactacttac
5521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
5581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
5641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
5701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
5761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
5821 gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
5881 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
5941 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
6001 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
6061 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
6121 ttaatagt
//
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