| 质粒名称: | pJKR-L-tetR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 62562 |
| 存储实验室: | George Church |
|
载体骨架基本信息 |
|
| 载体名称: | SC101 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 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: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pJKR-L-tetR was a gift from George Church (Addgene plasmid # 62562 ; http://n2t.net/addgene:62562 ; RRID:Addgene_62562) |
| 参考文献部分: | Synthetic biosensors for precise gene control and real-time monitoring of metabolites. Rogers JK, Guzman CD, Taylor ND, Raman S, Anderson K, Church GM. Nucleic Acids Res. 2015 Jul 7. pii: gkv616. 10.1093/nar/gkv616 PubMed 26152303 |
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 pJKR-L-tetR 5093 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Tetracycline biosensor. Low copy plasmid that expresses GFP
proportional to tetracycline concentration..
ACCESSION .
VERSION .
KEYWORDS pJKR-L-tetR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5093)
AUTHORS Rogers JK, Guzman CD, Taylor ND, Raman S, Anderson K, Church GM
TITLE Synthetic biosensors for precise gene control and real-time
monitoring of metabolites.
JOURNAL Nucleic Acids Res. 2015 Jul 7. pii: gkv616.
PUBMED 26152303
REFERENCE 2 (bases 1 to 5093)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5093
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 59..919
/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"
primer_bind complement(277..296)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2071..2293
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
primer_bind complement(2785..2802)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 2955..3041
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 3203..3221
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3228..3246
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 3313..4026
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
RBS 4319..4327
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 4335..4958
/codon_start=1
/gene="tetR"
/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 complement(4408..4427)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
terminator 5012..5059
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
661 ccggcaacaa ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggctctcg
781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
901 actgattaag cattggtaac tgtcatacca agtttactca tatatacttt agattgattt
961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
1021 caaaatccct taacgtggag ttttcgttcc actgagcgtc agaccccgaa cctcagatcc
1081 ttccgtgttt agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga
1141 acgaaccatt gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg
1201 tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt
1261 aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct
1321 caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag
1381 tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac
1441 ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca
1501 ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt
1561 gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact
1621 taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct
1681 cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga
1741 aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc
1801 tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt
1861 attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt
1921 tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac
1981 taatcgctag ttcatttgct ttgaaaacaa ctaatttaga catacatctc aattggtcta
2041 ggtgatttta atcactatac caattgagat gggctagtca atgataatta ctagtccttt
2101 tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt
2161 ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag
2221 tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc
2281 ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc
2341 tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa
2401 gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt
2461 ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg
2521 cactacaggc gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc
2581 gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt
2641 gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga
2701 caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa ctggctgttt
2761 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
2821 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
2881 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
2941 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
3001 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
3061 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
3121 atcaaattaa gcagaaggcc atccgtcagg atggcctttt tgcgtttcta caaactctgc
3181 tagcaagtaa ggccgactcg agtccctatc agtgatagag attgacatcc ctatcagtga
3241 tagagatact gagcacatca gcaggacgca tgaccgaatt cattaaattt aactttaaga
3301 aggagatata atatgcgtaa aggtgaagaa ctgttcaccg gtgttgttcc gatcctggtt
3361 gaactggacg gtgacgttaa cggtcacaaa ttctctgttc gtggtgaagg tgaaggtgac
3421 gctaccaacg gtaaactgac cctgaaattc atctgcacca ccggtaaact gccggttccg
3481 tggccgaccc tggttaccac cctgacctac ggtgttcagt gcttcgctcg ttacccggac
3541 cacatgaaac agcacgactt cttcaaatct gctatgccgg aaggttacgt tcaggaacgt
3601 accatctctt tcaaagacga cggtacctac aaaacccgtg ctgaagttaa attcgaaggt
3661 gacaccctgg ttaaccgtat cgaactgaaa ggtatcgact tcaaagaaga cggtaacatc
3721 ctgggtcaca aactggaata caacttcaac tctcacaacg tttacatcac cgctgacaaa
3781 cagaaaaacg gtatcaaagc taacttcaaa atccgtcaca acgttgaaga cggttctgtt
3841 cagctggctg accactacca gcagaacacc ccgatcggtg acggtccggt tctgttgccg
3901 gacaaccact acctgtctac ccagtctgtt ctgtctaaag acccgaacga aaaacgtgac
3961 cacatggttc tgctggaatt cgttaccgct gctggtatca cccacggtat ggacgaactg
4021 tacaaagatg catgccagtt ctagcataac cctaatgagt gagctaactt acattaattg
4081 cgttgcgcct taattaacgg cactcctcag caaatataat gaccctcttg ataacccaag
4141 agggcatttt ttaatgccca tggcgtttac cacagctaac accacgtcgt ccctatctgc
4201 tgccctaggt ctatgagtgg ttgctggata actttacggg catgcataag gctcgtaggc
4261 tatattcagg gagaccacaa cggtttccct ctacaaataa ttttgtttaa ctttgaaata
4321 aggaggtaat acaaatgtct cgtttagata aaagtaaagt gattaacagc gcattagagc
4381 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
4441 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
4501 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
4561 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
4621 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
4681 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
4741 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
4801 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
4861 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
4921 gattagaaaa acaacttaaa tgtgaaagtg ggtcttaata agcgactaaa aaattgaatg
4981 taggaaacca acatgccagt tcgagcaata actagcataa ccccttgggg cctctaaacg
5041 ggtcttgagg ggttttttgc tgaaacctca ggcgagaagc acacggtcac act
//
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