pCMV-Otogl(mouse)-3×FLAG-Neo
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Instructions
Ⅰ. Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature)
1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid;
2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min;
3. 42 degrees heat shock for 60s, ice bath for 2min, add 900μl of antibiotic-free LB medium, and culture at 180rpm shaking for 45min;
4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate;
5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation;
6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed.
Ⅱ. Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation).
1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall;
2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer.
Ⅲ. Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation).
1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall;
2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance;
3. Inverted culture in constant temperature incubator for 12~16h;
4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance;
5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.
Product Disclaimer
For Research Use Only research use only . They are not intended for clinical trials or for use in humans, including oral administration, injection, or therapeutic use in vivo or ex vivo.Plasmid Sequence and Experimental Performance reference/template plasmid . MiaoLing guarantees the accuracy of the specified gene sequence based on our sequencing results, but does not guarantee downstream experimental performance, including gene expression, transfection efficiency, protein expression, or other experimental outcomes.Sequence Verification of Certain Vector Products key functional regions has been confirmed to be correct, the product will be considered a correct and usable vector for research purposes.After-Sales Period for Small-Pack Plasmid Products three months from the date of receipt . Customers are advised to transform and process the plasmid promptly after receipt. After the three-month period, a new purchase may be required for further support or replacement.Research-Related Risks and Limitation of Liability Citation in Chinese Publications Citation in English Publications “Product Name (Product Item Number) was obtained from MiaoLing Plasmid, China.”
质粒序列
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GTTAGGCGTTTTGCGCTGCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCTGGCTAGCGTTTAAACTTAAGCTTGGTACCGAGCTCGGATCCACTAGTCCAGTGTGGTGGAATTCGCCACCatggtaccttggcgtgctctctcccttcctatactgctggtttcattacgaggatacgtttgtgcgtcatctgtcctgtcagaaacatcagaaagtgagttctatgaaaatgaacagagaagggctctgttagcagtacagtttgaagcaacctctccgagatattttttccacgaagctattaattggggtgagagtaaaataaaaggttcatgtcctcatgaatgccttaacggggctttctgttctaagacgggtacgtgtgactgtcaaatattccaggctctcgggacaaggtgccagattgtgccaaacatgggaagtggcagagatgggatttgcaaaacctggggacagtatcactttgaaacatttgatggaatctactattatttcccaggaagttgttcttatatatttgcaaaggactgtggtaatttggagcctcagtacacagtatgggtccataatagcccaaagtgccttggttccgtgtattcttgttaccgctccatcagcttatttttttcaaaccaagaggaaattagaatctatggacatgaaataagaaaaaatgggatcagtttatctctgcctcagacacttgggcaggtgcacttagagaaggtagctgactacattctggtgaagacgacctttggattttctttggcttgggatggaatatcgggaatttacctcaaactctcggaggagcacagagggaaatcatgtggcttgtgtgcaaattataatggcatccagtcagacgactttgtgatcctgcaagaggactacacagaagacattgctatgtttgcaaatagctggctggtgctaactccagatgatactaaatgtgtacctacgccctcagactttccaaatccatgttccagtggaatgccagcatttgaggcaatcttcttcaaatgccagatattgttacagttcccctttctgagctgccacgaatacattgatccatacttatacattgccagctgtgttaatgacctgtgcaaaactgatgacgatgaaacttactgtcgagcagccacagagtacgctagagcctgttcccacgccggcttccccattcaagactggagggatgactttccagcatgcactgataaatgtgacgatagcttcgtccaccgggactgtatcagttgttgccctccgagctgcacatttgagaagcagtgtcttgggagcaatcttcactgccttgatggatgctattgtgcagacggcctcataatggacaatggaacttgcatctctttggagagctgcccttgtagttttcacggattagcttattcagttggttcaaaaatcgaacaagagtgcactgaatgtgtatgtgttggtggagtttggaactgcaccgaacatgactgtccagttcagtgttctgttgtgggtgactctcatttcaccacctttgatgggcgtcattactcttttattggcttgtgtcagtacatccttgtgaaaggaactggaaaagatagattcacgatcactttgcagaaggctcactgtgaacagaacctgggtttggtctgccttcagtccataactctaatactggaggatgactttaacaaacaagtgacgctcagcagaggaggccagattgtcaccagtcccaaccaaggcttcaccctgaacggcattgttgaaattcaaaccctgtcatccttattcattctcctaagaaccacctttggtttaaagatcttgttcgctattgatggagaaagaatttacattcagctttccagtgcctggaaaagaaggacattaggtctgtgcggcacctttaatggcaacattagggatgattttctttctccatcaggcatgatcgaaggaaccccacagctccacgcacatgcgtggagagtctcctccacgtgtttcgcgcctgtccatgtcccgatggtggatccctgcaatattaaccaacagaacattgggtatgcagcacactgtgatgtcatccaccaggagctctttgctccttgccatgtctacgtcagtcccgggctgtactaccagttgtgtcgccacgatgcctgcaaatgtgggagcccctgcctgtgtaacgctctcgcccactacgcctacctgtgtggtcagcgtggtgtgcccattgatttcagagctcacatctccttctgtgctgtggtgtgtcagaagggcatgctgtaccaccactgctcgtcactctgcctccgctcctgcacctctctctcctcccctgagcagtgcaaagacgactgtgctgaagggtgcaactgtcctgaaggcaaattctatgaagagacacttaacttttgtgtgcccatatatcactgccggtgtcattatagggggagcatttaccagccaggggagctcattccaaccccctcaggcttatgccagtgttcaaatgggactgtgaaatgtgatgagctagcaacaccctctactgtccacgcctgcccagaagggaaggaatattttgactgcaggtttcctgaccctgcgttaccagctggtggtataaactgcgagacgacgtgcgcaaacctggccatgaacttcacctgtgctccatcctcgccttgcatcagtggctgcgtttgtgctgcaggaatggcagaacacaaaggaaaatgctatgttcccgaaagctgtccctgcatctggaaggactgggagtacagctcaggggaggtcatctctacaccgtgttacacctgtgtttgtcggcgggggatgttcaattgcacatattatccatgtcctgcagtatgcacagtatatggggaccggcattatcactcctttgatggattggaatatgactatatcagcgattgccaagtgtttttgataaagagtacagacgattctgatatatctgtgatttcccaaaacaagaaatgctttgacaatgatattgtttgttctaaaagtgttttgatatcaattggagacactgaaatatacctcaatgatgctccttacaaacagaagcgatcagggtttttcctggaaagcagacctgagtaccagctctggaaggctgggttctacatagtaatatacttcccggaggaagatatcaccattctgtgggatgagaagacaaccatccacatcaaagtcgggccacagtggaagaacaagctggcaggattgtgtggtaactttgacaaatgtacttccaatgacatgacaacatctaataacattgaagtgcgaaatgctcaggtgtttggagacagctgggccttaggacagtgtgaagatcttatggaggccctgaagccctgcgaggcacaccaaaacaaatttccctatgccaagagagaatgttccattttgtacagtgatgtttttgctccttgtcgcaatgtgattgacgtcacctcttttgccaagaactgccatgaagacacgtgtaactgtaacctgggtggggactgtgaatgcctatgtaccagtgtggccgcatacgcttacaaatgctgccaggaaggggtgcctgttcactggaggtcacccactgtctgtgcccttgactgtgaatattataatcaagggcttggggaaggaccttatatgctggcaagttatgggcaaagtggccttgttcttggtgccaatatgaccagtagaagcgttttctctttgcccagaagcaacaatcgtggtaatttattttttatttttatgatcactccgggccttttcaaagagaagacatcctcattggctcttgtttccttggaatctgctgaaaggcccaactactttctctatgtccacgacaatgacactcttagcttgaagctgtggcgggcaaactcagaatttcatcagagagcaacatttttccaccaccaaggcctctggattcctggctacagtgcattcgaattatacagcaagaaaggctacttcatcgtttttatgggttccagtgtcaaagcgtcaaaatacgatgactcggaagaattcaagcagtcaagtagctttagcatagaagaaatccaggcagtggtcccttacaggaggatgtgcgagtggagatacgaaccttgtgctactccgtgttttaaaacatgcagtgatccagaggccttagcctgtacattccttccaccggttgaaggatgcttgccatattgccccaagaatatgatcctggatgagacgactcttaaatgtgttcacccagaagactgcataccactgtttccaacagaaccagcattgccaccagacattaccccatcagacatgacaccgaccccaggtttggaatgcgagcctcaacaatttgatcctgtttataactgtagccaatatatctgccttaacatggagtggaccttctacaactggtctctgaattgcccgaaggacttggaaatgcccgactgtgggttccgaggctggcctgttcaagtgaatactgatatatgctgtccggagtgggaatgcccctgtcggtgctccatgttgtcagagttgagcatcattacatttgatgggaacagtgcagcactatctagtatggcttcttacatcttagtaagagtccccggggaaattgtagttgttcacattgacaaatgttccatgaatcagaatggacatgcccttaaaaaaccagcttcttttggacgaatctcaggactttgttttaagaagctaaatgtgacaacatctatacataaaatacttatcaacagagtggtaagaaaggtagacgtggattctattgttgtgcctttgcccttttcaagtcatgagctgttcatagaagattctggaacaatgtatgtgattactactcctgctggcttaatcataaagtgggctcaccttacaggaatcatagacattcacttcggcccccagtttaacttgtcatcttacacagaagggctttgtgggatttgcaatgacaatccagatgatgacctcaggatgcagaatggcaccatcattaccaacatggaagacatagagttatttattggcagctgggaaattgagaaatcatttgaagtgacaatgagaagacctgtgaggaactgtactgaatatgactgcagtcattgcatcgagttattgaacagagaaggtttcattccatgccatgacaaagtctcaccaagggacttttgtgaaaaaatgtggatcaactacacctacttctggagctacgaatgtgatgcaatctctgcatatgtggctttgtgcaacaaatttgatatctgcattcagtggcgcacccctgactattgccctctgagttgtccagagggaaaggaatatcagccctgtgtgagaccatgtgaagcaagaacatgtctcaacaaatggttctatggacactcctcatgtctgaatttgagagaagactgtgtgtgcaagaatggaaccattcttcaccggccagataaaacactatgcatcccagagcaagaatgtgtgtgcactgacagagaggagcacccccgcagtgctggggagatttggaatggtggcatagatgaatgcaccctctacaaatgtctggaggatggaaacattattcctatagagcctgtctgtgaagaagagccttcccccatctgtgaacgaacagctgaagtggtcataggcattgttgataagctgacctgctgttctaaaaaagtctgtggctgtgacatgagtttgtgtgacaggaccatcccaccatgcacaaatagccagaagttgatcgttggctatagtgctctttcttgttgcccacagtatgaatgtgaatgtgatacggtgagatgtcctgatatttcaacgccagtttgcagagatgaccagtttgtgcttcaagttcgacagggagagccttgctgtttctacccttcctgcgtttgtaagacatgcactgaaccaactccacaatgtaccgatggggaattcctcactgttaatatcaatactacacatttatgctgtcctcagtactactgcgtttgtgagccagatctttgccctccaccatcacttgagtgtgcaaaagatatgaacctggtgaaagaaaatgtatctggacaatgttgcccaaactggcgttgcgaatgcaactgtgagactcttgtcatgccaacctgtgacgtgggagaatttgctgcaatagatcaaaactttcagactgactgtggctgtgtacagtatctctgtgaaaaggatgatgtgtgtgtgttccaagaagtatcagtattgaatcctgggcaatctctaataaagtatttagaagaggaattttgttatataatcgaatgtctggatgaaaaagataactataccgactttcatactttgaatgttacaatggtgaactgttcaaaagactgtgatgctcaccagatatacattccatcctccagtgattatgattgctgtggaacatgtaaaaatatatcctgcaaatttattatggaaaatggaacatcagttatatatgaggaggggagcacctggcactataactgcagcacgtacgaatgtgttaacactgaggagggagcaacgattctaaactacagcatggtttgccctccttttaatgaaactgaatgcaaactgaatgaagggattgtgaagctttataatgaaggctgttgcaagatctgcaaacgagaagagcgaatatgccagaaagtgatcattaaatccatcataaagaaacaggactgtgtaagccaaagctctattagtgtggcatcttgtgatgggaaatgcccgtcagctaccatatataacatcaatgttgaaagtcacctgaggttctgcaagtgctgtcgtgaaaatggagtgcgaaacgtgacggtgcctctgcactgctcaggaaatggcactgaagtaatgtacacactccaggagccgatagactgcacatgccagtggaacCTCGAGGACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGACTACAAGGATGACGATGACAAGTAGTGAGGGCCCGATATCTCGACAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGGAAACGGGGGAGGCTAACTGAAACACGGAAGGAGACAATACCGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATACCCCACCGAGACCCCATTGGGGCCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCCACCCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAGGCCCTGCCATAGCAGATCTGCGCAGCTGGGGCTCTAGGGGGTATCCCCACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGCATCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGGGGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTAATTCTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCTGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAGCTCCCGGGAGCTTGTATATCCATTTTCGGATCTGATCAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAGCGGGACTCTGGGGTTCGCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCCAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGTATACCGTCGACCTCTAGCTAGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCGACGGATCGGGAGATCTCCCGATCCCCTATGGTCGACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGG
Plasmid Elements
CMV enhancer 50..429
/note="人类巨细胞病毒即刻早期增强子"
CMV promoter 430..633
/note="人巨细胞病毒（CMV）即刻早期启动子"
regulatory 700..709
/regulatory_class="other"
/note="脊椎动物强启动翻译的共识序列（Kozak，1987）"
Otogl-m 706..7680
/codon_start=1
/translation="MVPWRALSLPILLVSLRGYVCASSVLSETSESEFYENEQRRALLA
VQFEATSPRYFFHEAINWGESKIKGSCPHECLNGAFCSKTGTCDCQIFQALGTRCQIVP
NMGSGRDGICKTWGQYHFETFDGIYYYFPGSCSYIFAKDCGNLEPQYTVWVHNSPKCLG
SVYSCYRSISLFFSNQEEIRIYGHEIRKNGISLSLPQTLGQVHLEKVADYILVKTTFGF
SLAWDGISGIYLKLSEEHRGKSCGLCANYNGIQSDDFVILQEDYTEDIAMFANSWLVLT
PDDTKCVPTPSDFPNPCSSGMPAFEAIFFKCQILLQFPFLSCHEYIDPYLYIASCVNDL
CKTDDDETYCRAATEYARACSHAGFPIQDWRDDFPACTDKCDDSFVHRDCISCCPPSCT
FEKQCLGSNLHCLDGCYCADGLIMDNGTCISLESCPCSFHGLAYSVGSKIEQECTECVC
VGGVWNCTEHDCPVQCSVVGDSHFTTFDGRHYSFIGLCQYILVKGTGKDRFTITLQKAH
CEQNLGLVCLQSITLILEDDFNKQVTLSRGGQIVTSPNQGFTLNGIVEIQTLSSLFILL
RTTFGLKILFAIDGERIYIQLSSAWKRRTLGLCGTFNGNIRDDFLSPSGMIEGTPQLHA
HAWRVSSTCFAPVHVPMVDPCNINQQNIGYAAHCDVIHQELFAPCHVYVSPGLYYQLCR
HDACKCGSPCLCNALAHYAYLCGQRGVPIDFRAHISFCAVVCQKGMLYHHCSSLCLRSC
TSLSSPEQCKDDCAEGCNCPEGKFYEETLNFCVPIYHCRCHYRGSIYQPGELIPTPSGL
CQCSNGTVKCDELATPSTVHACPEGKEYFDCRFPDPALPAGGINCETTCANLAMNFTCA
PSSPCISGCVCAAGMAEHKGKCYVPESCPCIWKDWEYSSGEVISTPCYTCVCRRGMFNC
TYYPCPAVCTVYGDRHYHSFDGLEYDYISDCQVFLIKSTDDSDISVISQNKKCFDNDIV
CSKSVLISIGDTEIYLNDAPYKQKRSGFFLESRPEYQLWKAGFYIVIYFPEEDITILWD
EKTTIHIKVGPQWKNKLAGLCGNFDKCTSNDMTTSNNIEVRNAQVFGDSWALGQCEDLM
EALKPCEAHQNKFPYAKRECSILYSDVFAPCRNVIDVTSFAKNCHEDTCNCNLGGDCEC
LCTSVAAYAYKCCQEGVPVHWRSPTVCALDCEYYNQGLGEGPYMLASYGQSGLVLGANM
TSRSVFSLPRSNNRGNLFFIFMITPGLFKEKTSSLALVSLESAERPNYFLYVHDNDTLS
LKLWRANSEFHQRATFFHHQGLWIPGYSAFELYSKKGYFIVFMGSSVKASKYDDSEEFK
QSSSFSIEEIQAVVPYRRMCEWRYEPCATPCFKTCSDPEALACTFLPPVEGCLPYCPKN
MILDETTLKCVHPEDCIPLFPTEPALPPDITPSDMTPTPGLECEPQQFDPVYNCSQYIC
LNMEWTFYNWSLNCPKDLEMPDCGFRGWPVQVNTDICCPEWECPCRCSMLSELSIITFD
GNSAALSSMASYILVRVPGEIVVVHIDKCSMNQNGHALKKPASFGRISGLCFKKLNVTT
SIHKILINRVVRKVDVDSIVVPLPFSSHELFIEDSGTMYVITTPAGLIIKWAHLTGIID
IHFGPQFNLSSYTEGLCGICNDNPDDDLRMQNGTIITNMEDIELFIGSWEIEKSFEVTM
RRPVRNCTEYDCSHCIELLNREGFIPCHDKVSPRDFCEKMWINYTYFWSYECDAISAYV
ALCNKFDICIQWRTPDYCPLSCPEGKEYQPCVRPCEARTCLNKWFYGHSSCLNLREDCV
CKNGTILHRPDKTLCIPEQECVCTDREEHPRSAGEIWNGGIDECTLYKCLEDGNIIPIE
PVCEEEPSPICERTAEVVIGIVDKLTCCSKKVCGCDMSLCDRTIPPCTNSQKLIVGYSA
LSCCPQYECECDTVRCPDISTPVCRDDQFVLQVRQGEPCCFYPSCVCKTCTEPTPQCTD
GEFLTVNINTTHLCCPQYYCVCEPDLCPPPSLECAKDMNLVKENVSGQCCPNWRCECNC
ETLVMPTCDVGEFAAIDQNFQTDCGCVQYLCEKDDVCVFQEVSVLNPGQSLIKYLEEEF
CYIIECLDEKDNYTDFHTLNVTMVNCSKDCDAHQIYIPSSSDYDCCGTCKNISCKFIME
NGTSVIYEEGSTWHYNCSTYECVNTEEGATILNYSMVCPPFNETECKLNEGIVKLYNEG
CCKICKREERICQKVIIKSIIKKQDCVSQSSISVASCDGKCPSATIYNINVESHLRFCK
CCRENGVRNVTVPLHCSGNGTEVMYTLQEPIDCTCQWN"
CDS 7687..7752
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
Cleavage site after base 7752"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
WPRE 7776..8364
/note="土拨鼠肝炎病毒转录后调控元件"
HSV TK poly(A) signal 8430..8478
/note="单纯疱疹病毒胸苷激酶多聚腺苷酸化信号（Cole和Stacy，1985）"
f1 ori 8680..9108
/direction=RIGHT
/note="f1噬菌体复制起点；箭头表示（+）链合成的方向"
SV40 promoter 9122..9451
/note="SV40增强子和早期启动子"
SV40 ori 9302..9437
/note="SV40复制起点"
NeoR/KanR 9518..10312
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="赋予对新霉素、卡那霉素和G418（遗传霉素（R））的抗性"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
SV40 poly(A) signal 10488..10609
/note="SV40聚腺苷酸化信号"
primer_bind complement(10658..10674)
/note="常见的测序引物，多种相似变体之一"
lac operator 10682..10698
/bound_moiety="lac repressor encoded by lacI"
/note="lac阻遏物与lac操作员结合，抑制大肠杆菌中的转录。这种抑制可以通过添加乳糖或异丙基β-D-硫代吡喃半乳糖苷（IPTG）来缓解。"
lac promoter complement(10706..10736)
/note="大肠杆菌lac操纵子的启动子"
CAP binding site 10751..10772
/bound_moiety="E. coli catabolite activator protein"
/note="CAP结合在cAMP存在下激活转录。"
ori complement(11060..11648)
/direction=LEFT
/note="高拷贝数ColE1/pMB1/pBR322/pUC复制起点"
AmpR complement(11819..12679)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="对氨苄青霉素、羧苄青霉素和相关抗生素产生耐药性"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
AmpR promoter complement(12680..12784)
/gene="bla"
-3×FLAG-Neo 图谱.png)
