Difference between revisions of "Part:BBa K2148001"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | The aadA gene confers antibiotic resistance to Spectinomycin and Streptomycin. | + | The aadA gene confers antibiotic resistance to Spectinomycin and Streptomycin and is functional in both chloroplasts and <i>E. coli</i>. |
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| + | This part can be used with other level 0 Phytobrick to create a transcriptional unit to provide a selection pressure on transformations. | ||
| + | |||
| + | Previous experience with this antibiotic marker in Saul Purton's lab recommends the psaA promoter, which is a strong promoter. However, when constructing level 2 composite parts, it is advisable to use the psaA promoter for the gene of interest and place aadA under atpA promoter which is weaker. | ||
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| + | ===Verification=== | ||
| + | |||
| + | Part verified in <i>E. coli</i> ligated into Paul Surton's (UCL) pASap1 backbone. These transformed bacteria grow on both ampicilin (backbone antibioti) and spectinomycin (albeit slowly). Picture below indicates growth on Spec plates (before plating on amplicilin plates to verify double resistance). | ||
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| + | <b>(PUT VERIFIED PHOTO HERE)</b> | ||
| + | |||
| + | |||
| + | |||
| + | ===Sequence=== | ||
| + | |||
| + | Sequencing data obtained from Biosource sequencing | ||
| + | ATTTNACGTAGTGGACAAATTCTTCCNACTGATCTGCGCGCGAGGCCAAGCGATCTTCTTCTTGTCCAAGATAAGCCTGTCTAGC | ||
| + | TTCAAGTATGACGGGCTGATACTGGGCCGGCAGGCGCTCCATTGCCCAGTCGGCAGCGACATCCTTCGGCGCGATTTTGCCGGTT | ||
| + | ACTGCGCTGTACCAAATGCGGGACAACGTAAGCACTACATTTCGCTCATCGCCAGCCCAGTCGGGCGGCGAGTTCCATAGCGTTA | ||
| + | AGGTTTCATTTAGCGCCTCAAATAGATCCTGTTCAGGAACCGGATCAAAGAGTTCCTCCGCCGCTGGACCTACCAAGGCAACGCT | ||
| + | ATGTTCTCTTGCTTTTGTCAGCAAGATAGCCAGATCAATGTCGATCGTGGCTGGCTCGAAGATACCTGCAAGAATGTCATTGCGC | ||
| + | TGCCATTCTCCAAATTGCAGTTCGCGCTTANCTGGATAACGCCACGGAATGATGTCGTCGTGCACAACAATGGTGACTTCTACAG | ||
| + | CGCGGAGAATCTCGCTCTCTCCAGGGGAAGCCGAAGTTTCCAAAANGTCGTTGATCAAAGCTCGCCGCGTTGTTTCATCAAGCCT | ||
| + | TANGTCACCGTAACCAGCAGATCAATATCACTGTGTGGCTTCNNCCGCCATCCACTGCGGAGCCGTACAAATGTACGGCCAGCAA | ||
| + | CGTCNTTCGAGATGGCGCTCGATGACGCCAACTACCTCTGATANTTGAGTTGANACTTCGGCGATACCGCTTCACGANCCATTCG | ||
| + | AGACCACTCNTCGCTNCTAAANANNGGCCNCAATTCCNNNNNTCNNNNNNNCCNAANGCTAAGGATTTTNTTAATCTGAAATTCN | ||
| + | NGCCTNNNGATACNCTAATTTTANAGGTAATGNCANNNNNANNNNNCNNNNANN | ||
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<partinfo>BBa_K2148001 parameters</partinfo> | <partinfo>BBa_K2148001 parameters</partinfo> | ||
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| + | ===References=== | ||
| + | Goldschimdt-Clermont, M.: Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker of site-directed transformation of chlamydomonas. Nucleic Acids Res. 1991 Aug 11; 19(15): 4083–4089. Online at: | ||
| + | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC328544/ | ||
===Characterization=== | ===Characterization=== | ||
Revision as of 16:24, 13 October 2016
aadA
A commonly used antibiotic for Chlamydomonas chloroplasts. Confers resistance to spectinomycin and streptomycin
Usage and Biology
The aadA gene confers antibiotic resistance to Spectinomycin and Streptomycin and is functional in both chloroplasts and E. coli.
This part can be used with other level 0 Phytobrick to create a transcriptional unit to provide a selection pressure on transformations.
Previous experience with this antibiotic marker in Saul Purton's lab recommends the psaA promoter, which is a strong promoter. However, when constructing level 2 composite parts, it is advisable to use the psaA promoter for the gene of interest and place aadA under atpA promoter which is weaker.
Verification
Part verified in E. coli ligated into Paul Surton's (UCL) pASap1 backbone. These transformed bacteria grow on both ampicilin (backbone antibioti) and spectinomycin (albeit slowly). Picture below indicates growth on Spec plates (before plating on amplicilin plates to verify double resistance).
(PUT VERIFIED PHOTO HERE)
Sequence
Sequencing data obtained from Biosource sequencing ATTTNACGTAGTGGACAAATTCTTCCNACTGATCTGCGCGCGAGGCCAAGCGATCTTCTTCTTGTCCAAGATAAGCCTGTCTAGC TTCAAGTATGACGGGCTGATACTGGGCCGGCAGGCGCTCCATTGCCCAGTCGGCAGCGACATCCTTCGGCGCGATTTTGCCGGTT ACTGCGCTGTACCAAATGCGGGACAACGTAAGCACTACATTTCGCTCATCGCCAGCCCAGTCGGGCGGCGAGTTCCATAGCGTTA AGGTTTCATTTAGCGCCTCAAATAGATCCTGTTCAGGAACCGGATCAAAGAGTTCCTCCGCCGCTGGACCTACCAAGGCAACGCT ATGTTCTCTTGCTTTTGTCAGCAAGATAGCCAGATCAATGTCGATCGTGGCTGGCTCGAAGATACCTGCAAGAATGTCATTGCGC TGCCATTCTCCAAATTGCAGTTCGCGCTTANCTGGATAACGCCACGGAATGATGTCGTCGTGCACAACAATGGTGACTTCTACAG CGCGGAGAATCTCGCTCTCTCCAGGGGAAGCCGAAGTTTCCAAAANGTCGTTGATCAAAGCTCGCCGCGTTGTTTCATCAAGCCT TANGTCACCGTAACCAGCAGATCAATATCACTGTGTGGCTTCNNCCGCCATCCACTGCGGAGCCGTACAAATGTACGGCCAGCAA CGTCNTTCGAGATGGCGCTCGATGACGCCAACTACCTCTGATANTTGAGTTGANACTTCGGCGATACCGCTTCACGANCCATTCG AGACCACTCNTCGCTNCTAAANANNGGCCNCAATTCCNNNNNTCNNNNNNNCCNAANGCTAAGGATTTTNTTAATCTGAAATTCN NGCCTNNNGATACNCTAATTTTANAGGTAATGNCANNNNNANNNNNCNNNNANN
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 652
- 1000COMPATIBLE WITH RFC[1000]
References
Goldschimdt-Clermont, M.: Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker of site-directed transformation of chlamydomonas. Nucleic Acids Res. 1991 Aug 11; 19(15): 4083–4089. Online at: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC328544/
Characterization
References
Goldschmidt-Clermont M. Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker of site-directed transformation of chlamydomonas. Nucleic Acids Research. 1991;19(15):4083-4089.