| 质粒名称: | pM1s3AsR_TS-C7 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 137945 |
| 存储实验室: | Neel Joshi |
|
载体骨架基本信息 |
|
| 载体名称: | pMUT1 |
| 载体出品公司: | Native to E. coli Nissle |
| 空载体大小: | 3173 bp |
| 载体修饰: | pMUT1用带有选择标记的绝缘盒修饰 |
| 载体类型: | 细菌表达，合成生物学；用于小鼠肠道中的大肠杆菌Nissle |
| 载体抗性: | 氨苄霉素,100 μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | Mach1 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mRFP1 |
| 物种来源: | Synthetic |
| 插入大小: | 678 bp |
| 启动子: | J23101 |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | GTTCCTTATCATCTGGCGAATCGG |
| 3’测序引物 | GAGACGAGACGAGACAGCCTG |
|
文献引用方法 |
|
| 材料和方法部分: | pM1s3AsR_TS-C7 was a gift from Neel Joshi (Addgene plasmid # 137945 ; http://n2t.net/addgene:137945 ; RRID:Addgene_137945) |
| 参考文献部分: | Plasmid Vectors for in Vivo Selection-Free Use with the Probiotic E. coli Nissle 1917. Kan A, Gelfat I, Emani S, Praveschotinunt P, Joshi NS. ACS Synth Biol. 2020 Dec 10. doi: 10.1021/acssynbio.0c00466. 10.1021/acssynbio.0c00466 PubMed 33301298 |
买家导航
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 pM1s3AsR_TS-C7 8584 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION E. coli Nissle pMUT1-derived plasmid with ampicillin resistance and
constitutive RFP expression and strong temperature sensitive GFP
expression.
ACCESSION .
VERSION .
KEYWORDS pM1s3AsR_TS-C7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8584)
AUTHORS Kan A, Gelfat I, Emani S, Praveschotinunt P, Joshi NS
TITLE Plasmid Vectors for in Vivo Selection-Free Use with the Probiotic E.
coli Nissle 1917.
JOURNAL ACS Synth Biol. 2020 Dec 10. doi: 10.1021/acssynbio.0c00466.
PUBMED 33301298
REFERENCE 2 (bases 1 to 8584)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8584
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 170..199
/label=T3Te terminator
/note="phage T3 early transcription terminator"
misc_feature 246..267
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
CDS 347..1207
/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"
terminator 1337..1366
/label=T3Te terminator
/note="phage T3 early transcription terminator"
RBS 1491..1502
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1509..2186
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 2231..2302
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2318..2345
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_feature 2352..2372
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 2408..2439
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
misc_RNA 2784..2833
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
CDS 2888..2905
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2918..3631
/codon_start=1
/product="green variant of teal fluorescent protein mTFP1
(Ai et al., 2008)"
/label=mWasabi
/translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
TNTINLEVKEGAPLPFSYDILTTAFSYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK
GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA
VARNSTDGMDELYKG"
promoter 3685..3762
/gene="lacI"
/label=lacI promoter
CDS 4918..4962
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
terminator 5014..5061
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS 5233..5250
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 5317..5364
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 7265..7284
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7507..7524
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 ctaaatctag aatgcggtcg agctcgtaaa cttggtctga cagctctagc tccggcaaaa
61 aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg
121 tttgccacct gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc
181 ccacccgtga aggtgagcca gtgagttgat tgctacgtaa ttagttagtt agcccttagt
241 gactcgaatt cgcggccgct tctagagcta aatacattca aatatctatc cgctcatgag
301 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga gtattcaaca
361 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
421 agaaacgctg gtgaaagtaa aagatgccga agatcagttg ggtgcacgtg tgggttacat
481 cgaactggac ctcaacagcg gtaagattct tgagagtttt cgccccgaag aacgtttccc
541 aatgatgagc acttttaaag ttctgctctg tggcgcggta ttatcccgta ttgacgccgg
601 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
661 agtcacagaa aagcatctta cggacggcat gacagtacgc gaattatgca gcgctgccat
721 aaccatgagt gataacacgg cggccaactt acttctgaca acgatcggag gaccgaagga
781 gcttaccgct tttttgcaca acatgggtga tcatgtaact cgccttgatc gttgggaacc
841 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagctatggc
901 aacaacgttg cgcaaactct taactggcga acttcttact ctcgcttccc ggcaacaatt
961 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
1021 tggctggttt attgctgata aatctggagc cggtgagcgt gggtcccgcg gtattattgc
1081 agccctgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagcca
1141 ggcaactatg gacgaacgta atcgccagat cgctgagata ggtgcctccc tgattaagca
1201 ttggtaataa tactagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt
1261 ctttaaaacc gaaaagatta cttcgcgttt gccacctgac gtctaagaaa aggaatattc
1321 agcaatttgc ccgtgccgaa gaaaggccca cccgtgaagg tgagccagtg agttgattgc
1381 tacgtaacaa tacgcaaacc gcctctcgtt ccttatcatc tggcgaatcg gacccacaag
1441 agcactgttt acagctagct cagtcctagg tattatgcta gctactagag aaagaggaga
1501 aatactagat ggcttcctcc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
1561 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
1621 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
1681 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
1741 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
1801 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
1861 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
1921 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
1981 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
2041 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
2101 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
2161 aaggtcgtca ctccaccggt gcttaataac gctgatagtg ctagtgtaga tcgctactag
2221 agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
2281 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
2341 ctgcgtttat atactagtag cggccgctgc aggagtcact aagggttagt tagttagatt
2401 agcagaaagt caaaagcctc cgaccggagg cttttgacta aaacttccct tggggttatc
2461 attggggctc actcaaaggc ggtaatcaga tgcggcgagc ggtatcagct cactcaaagg
2521 cggtaatcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgc tagtacgtac
2581 ggtttcgaag cttgctcttc ttaagcatta ctcgcatcca ttctcaggct gtctcgtctc
2641 gtctcctgac ctcctgccag caatagtaag acaacacgca aagtccgatc tcgatcccgc
2701 gaaattttaa tttgtttgtt agttagttta tttgttggtt tgtttgtgtt atacaataag
2761 cctctagaaa taattttgtt tagctgtcac cggatgtgct ttccggtctg atgagtccgt
2821 gaggacgaaa cagcctctac aaattttgtt taaaacttta agaaggagat ataccatggg
2881 cagcagccat caccatcatc accacagcca ggatccaatg gtgagcaagg gcgaggagac
2941 cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg gcaacgtgaa
3001 tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg gcaccaacac
3061 catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca ttctgaccac
3121 cgcgttcagt tacggcaaca gggccttcac caagtacccc gacgacatcc ccaactactt
3181 caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg aggacaaggg
3241 catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct acgagataca
3301 cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaaggaga ccaccggctg
3361 ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg acgtcaagat
3421 gaagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca tctacagggc
3481 caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg agatcctgaa
3541 ccacgacaag gactacaaca aggtgaccgt ttacgagatc gccgtggccc gcaactccac
3601 cgacggcatg gacgagctgt acaagggctg aaagcttgcg gccgcataat gcttaagtcg
3661 aacagaaagt aatcgtatga gctcgacacc atcgaatggc gcaaaacctt tcgcggtatg
3721 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtataaga aggagatata
3781 catatgcgtc cggcgacata cgaaccagaa cagattattg aagcagggct ggccctgcag
3841 gctgaaggac ggaatatcac cgggttcgca ctacgtaacc aggtgggtgg cggcaatccg
3901 acacgtctcc gccagatatg ggacgaatac caggcttcac agagcacggt cgtcactgaa
3961 ctcgttgccg agctgccagt ggaagtggct gaagaagtga aggccgtctc cgccgcgctg
4021 tccgaacgca tcacccagct ggcgacagaa ctgaatgaca aggcggtccg ggctgcagaa
4081 cgccgggttg cggaagtcac gcgtgctgcc ggtgaacaga ccgcacaggc agagcgggag
4141 ctggccgacg ccgcgcagac agtcgacgac ctggaagaaa aactggttga actgcaggac
4201 agatatgaca gtttgacgct ggcgctggag tcagaacgtt cactgcgtca gcagcatgat
4261 gtggagatgg cccagctgaa agagcgtctt gcggccgctg aagagaatac ccgtcagcga
4321 gaggaacggt atcaggagca gaggacagtg ctgcaggatg cgcttaatgc ggagcaggca
4381 cagcacataa acacgcggga agaccagcag aaacgactgg agcaaatttc tgccgaagct
4441 aatgcgcgta cagaagaact gaagtctgaa cgcgataaag tcaatactct ccttacccgc
4501 cttgaatcgc aggaaaatgc gctggcctca gaacgtcagc agcatctggc cacccgcgaa
4561 acgctgcagc aacgcctcga gcaggccatc gctgacacgc aggcgcgcgc cggtgagatt
4621 gcacttgaac gtgacagagt cagcagcctc accgcaaggc tggaatcgca ggaaaaggcc
4681 tcctcggagc aactggtgcg tatgggcagt gaaatagcca gtctgacaga gcgttgcaca
4741 cagctggaaa accagcgtga tgatgcccgt ctggagacga tgggggagaa agaaacggtc
4801 gcggcactgc gtggtgaggc tgaagccctg aagcgtcaga accagtcact gatggcggcg
4861 ctttcaggca ataaacagac cggtggccag aatgcgtgag gtaccctcga gtctggtaaa
4921 gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa
4981 ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
5041 cgggtcttga ggggtttttt gctgactcgg taccaaattt tcgaaaaaag acgctgaaaa
5101 gcgtcttttt tcgttttggt ccgagccaac tccctttaca acctcactca agtccgttag
5161 agccaggata catagattac cacaactccg agcccttcca cccgccggcg agacctctag
5221 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
5281 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
5341 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata aaccttgctt
5401 cttcgcggtg tttcagtggt gcgtacaatt aagggattat ggtaaatata tgagctatgc
5461 gataacttta actgtgaagc gatgaaccca ttacaggcaa agccaattac tcctgacagt
5521 ggtttagcca gaagcagggc taccaagacc aatgcaataa gtaatatatc gttttgctat
5581 cgtgcatcgc tcgcgctcag ttccattgtg cttttttaag ctgtcgtttt tcttacggta
5641 tataccggtt ttttatggcg tggtttctta acttgttcag ctactgatgg acccattgta
5701 tctaggtagt caactagctt tgttagatca taaatattgc gaccaccata tcggcgatca
5761 ctcttcgatg ttggtttctt gtccaagaga ttagcttttt caagatcatt gatagctctc
5821 tgaacagtcc gtacagaaac ccccatacgt atggctagac tttccattga cggatgcggc
5881 cactcttgca aactccacca gtgaacgatc aggttaagta gtgtgttaaa ggccactgaa
5941 gttagctttt tctcgttttg tataaaaaac aatacggtag gcactgctgt ccagccaaga
6001 gacaaaccgc cagctttcca tttattctta acggagtaag tcattgattt tcctaagccc
6061 caaaatattt aaagtatata ttatatggta tattcatatg aatagggtga cactggcgcc
6121 attattgtgc aaccaaaaaa gactactctg aaaacgagga aaagattttt tcctgcctga
6181 attagatacg gagttagcga tatgaaaacc gaacaacgtc atgatcttgt taaagatatt
6241 gaggtttttg gcgtatcctt gtctctgttg atttccagag cgaatgagaa gtctgttaca
6301 atgccatctg gtctaagtca ggagcagaga agagcatggg cagcggagca ggcgcgcaaa
6361 atccacaatt gaatattgtc tcattctctg agaccttcaa cctttattac acatccagat
6421 attctgcaaa aacactcgat aaaatcgatg atttcattga gcattttgaa aaatacaatc
6481 tctttggcga tcctttaaaa ggatatccag cttggactgg caaagtatcg ccatcgtgga
6541 aagtgcctga tcattacgaa aacaaagaag ctattgagaa gtatgctaga gctaacaaat
6601 tatggcatgc tcatttaggc gatccggttt ttaaagatac gtttcatggg aaatacaagg
6661 tttctgactg ggttattcat ttccagcggc tgacaccgaa ccatataaag cttttagagc
6721 ttggatacca tgacccaatg aagctacctc aaaactttga atgatcgagc ggcaggctaa
6781 atgaaatctt gagattcatt cagtctcgtc gtaatctcac tattgtaaaa acgaaaaaac
6841 caccctggca ggtggttttt cgaaggttag ttaatcctgg cagattctct aaccgtggta
6901 acagtcttgt gcgagacatg tcaccaaata ctgtcctttc agtgtagcct cagttaggcc
6961 gccacttcaa gaactctcgt tacatctctc gcacatcctg cttaccagtg gccgttgcca
7021 gtggcgttaa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
7081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
7141 ccgaaccgag ataccaacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
7201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
7261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
7321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc tcccgcgcgc
7381 ccttcttctg ggtctttgtc ttttgctcac atgttctttc cggttttatc ccccgattct
7441 gtggataacc gtattaccgc ctttgagtga gctgacaccg ctcgccgcag tcgaacgacc
7501 gagcgtagcg agtcagtgag cgaggaagcg gaagagcgcc ttatgtgaca ttttctcctt
7561 acgctcctct atgccgttct gcatcctgtc cggatgcgtt atatcccggt aagattttcc
7621 gcttcaaagc gtgtctgtat gctgttctgg agttcttctg cgagttcgtg cagtttctca
7681 cacatggcgg cctgttcgtc ggcattgagt gcgtccagtt tttcgagcag cgtcaggctc
7741 tgacttttta tgaatcccgc catgttgagt acggcttgct gctgcttatt catcttttcg
7801 ttttctccgt tctgtctgtc atctgcgttg tgtgattata tcgcgtacca cttttcgact
7861 gttttgctgc cgctattctg ccgcttggct ttttgacggg catttctgtc agacaacact
7921 gtcactgcca aaaaactgcc gtgcctttgt cggtaattcg agcttgctga caggacagga
7981 tgtgcaattg ttataccgcg catacatgca cgctattaca attgccctgg tcagggcttt
8041 gccccgacac ccatgtcaga tacggagcca tgttttatga caaaacgaag tggaagtaat
8101 acgcgcaggc gggctatcag tcgccctgtt cgtctgacgg cagaagaaga ccaggaaatc
8161 agaaaaaggg ctgctgaatg cggcaagacc gtttccggtt ttttacgggc ggcagctctc
8221 ggtaagaaag ttaactcact gactgatgat cgggtactga aagaagttat gagactgggg
8281 gcgttacaga aaaaactctt tatcgacggc aagcgtgtcg gggacaggga gtatgcggag
8341 gtgctgatcg ctattacgga gtatcaccgt gccctgttat ccaggcttat ggcagattag
8401 cttcccggag agaaaactgt cgaaaactga cggtatgaac accgtaagct cccaaagtga
8461 tcaccattcg ctttcatgca tagctatgca gcgagctgaa aacgatcctg acgcatcctt
8521 cctgtttttc cggggtaaaa catctctttt tgcggtgtct gcgtcagaat cgcgttcagc
8581 gccc
//
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