pDSK-GFPuv
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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
View Plasmid Sequence
TATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGAGCAAGACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATGATGATATATTTTTATCTTGTGCAATGTAACATCAGAGATTTTGAGACACAACGTGGCTTTGTTGAATAAATCGAACTTTTGCTGAGTTGAAGGATCAGATCACGCATCTTCCCGACAACGCAGACCGTTCCGTGGCAAAGCAAAAGTTCAAAATCACCAACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCTATTCTGACCTTGCCATCACGACTGTGCTGGTCATTAAACGCGTATTCAGGCTGACCCTGCGCGCTGCGCAGGGCTTTATTGATTCCATTTTTACACTGATGAATGTTCCGTTGCGCTGCCCGGATTACAGCTGCGTTTCCTGTTTGGGGTCGTTTGCGGGAAGGGGCGGAATCCTACGCTAAGGCTTTGGCCAGCGATATTCTCCGGTGAGATTGATGTGTTCCCAGGGGATAGGAGAAGTCGCTTGATATCTAGTATGACGTCTGTCGCACCTGCTTGATCGCGGCCGCGATAGCTAGATCGCGTTGCTCCTCTTCTCCATCCGCGTTCCAAGCTGCGGAAAGGCACCCATAAGCGTACGCCTGGTCGAGCAGGCGACGCGGATCGACGTCCAGCGCACGAGAGAATGCGTCCGCCATCTGTGCAATGCGTCTAGGATCGAGACAAAGGTCGTCTCTGTCAGCCGGATCGTAGAACATATTGGCGGCGCCAAAGCCCACTTCACCGACCAGACATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTCGAGCTCGGTACCCGGGGATCTAGAGGATCTCAATGAATATTGGTTGACACGGGCGTATAAGACATGTTATACTGTTGAATAACAAGCTTTCAATTCTCTATTTTGAGAAGAATTAATGTGCTTGGGAGTCCCTGATGATTAAATTAAATAAACCAAGATTTTACCATGACTGCAATTTTAGAGAGACGGGGGATCGATCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGGTGTTCAATGCTTTTCCCGTTATCCGGATCATATGAAACGGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAACGCACTATATCTTTCAAAGATGACGGGAACTACAAGACGCGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATCGTATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTCGGACACAAACTCGAGTACAACTATAACTCACACAATGTATACATCACGGCAGACAAACAAAAGAATGGAATCAAAGCTAACTTCAAAATTCGCCACAACATTGAAGATGGATCAGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCGACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGCGTGACCACATGGTCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAGCTCTACAAATAAGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAAGAAGAGCATACATCTGGAAGCAAAGCCAGGAAAGCGGCCTATGGAGCTGTGCGGCAGCGCTCAGTAGGCAATTTTTCAAAATATTGTTAAGCCTTTTCTGAGCATGGTATTTTTCATGGTATTACCAATTAGCAGGAAAATAAGCCATTGAATATAAAAGATAAAAATGTCTTGTTTACAATAGAGTGGGGGGGGTCAGCCTGCCGCCTTGGGCCGGGTGATGTCGTACTTGCCCGCCGCGAACTCGGTTACCGTCCAGCCCAGCGCGACCAGCTCCGGCAACGCCTCGCGCACCCGCTGGCGGCGCTTGCGCATGGTCGAACCACTGGCCTCTGACGGCCAGACATAGCCGCACAAGGTATCTATGGAAGCCTTGCCGGTTTTGCCGGGGTCGATCCAGCCACACAGCCGCTGGTGCAGCAGGCGGGCGGTTTCGCTGTCCAGCGCCCGCACCTCGTCCATGCTGATGCGCACATGCTGGCCGCCACCCATGACGGCCTGCGCGATCAAGGGGTTCAGGGCCACGTACAGGCGCCCGTCCGCCTCGTCGCTGGCGTACTCCGACAGCAGCCGAAACCCCTGCCGCTTGCGGCCATTCTGGGCGATGATGGATACCTTCCAAAGGCGCTCGATGCAGTCCTGTATGTGCTTGAGCGCCCCACCACTATCGACCTCTGCCCCGATTTCCTTTGCCAGCGCCCGATAGCTACCTTTGACCACCATGGCATCAGCGGTGACGGCCTCCCACTTGGGTTCCAGGAACAGCCGGAGCTGCCGTCCGCCTTCGGTCTTGGGTTCCGGGCCAAGCACTAGGCCATTAGGCCCAGCCATGGCCACCAGCCCTTGCAGGATGCGCAGATCATCAGCGCCCAGCGGCTCCGGGCCGCTGAACTCGATCCGCTTGCCGTCGCCGTAGTCATACGTCACGTCCAGCTTGCTGCGCTTGCGCTCGCCCCGCTTGAGGGCACGGAACAGGCCGGGGGCCAGACAGTGCGCCGGGTCGTGCCGGACGTGGCTGAGGCTGTGCTTGTTCTTAGGCTTCACCACGGGGCACCCCCTTGCTCTTGCGCTGCCTCTCCAGCACGGCGGGCTTGAGCACCCCGCCGTCATGCCGCCTGAACCACCGATCAGCGAACGGTGCGCCATAGTTGGCCTTGCTCACACCGAAGCGGACGAAGAACCGGCGCTGGTCGTCGTCCACACCCCATTCCTCGGCCTCGGCGCTGGTCATGCTCGACAGGTAGGACTGCCAGCGGATGTTATCGACCAGTACCGAGCTGCCCCGGCTGGCCTGCTGCTGGTCGCCTGCGCCCATCATGGCCGCGCCCTTGCTGGCATGGTGCAGGAACACGATAGAGCACCCGGTATCGGCGGCGATGGCCTCCATGCGACCGATGACCTGGGCCATGGGGCCGCTGGCGTTTTCTTCCTCGATGTGGAACCGGCGCAGCGTGTCCAGCACCATCAGGCGGCGGCCCTCGGCGGCGCGCTTGAGGCCGTCGAACCACTCCGGGGCCATGATGTTGGGCAGGCTGCCGATCAGCGGCTGGATCAGCAGGCCGTCAGCCACGGCTTGCCGTTCCTCGGCGCTGAGGTGCGCCCCAAGGGCGTGCAGGCGGTGATGAATGGCGGTGGGCGGGTCTTCGGCGGGCAGGTAGATCACCGGGCCGGTGGGCAGTTCGCCCACCTCCAGCAGATCCGGCCCGCCTGCAATCTGTGCGGCCAGTTGCAGGGCCAGCATGGATTTACCGGCACCACCGGGCGACACCAGCGCCCCGACCGTACCGGCCACCATGTTGGGCAAAACGTAGTCCAGCGGTGGCGGCGCTGCTGCGAACGCCTCCAGAATATTGATAGGCTTATGGGTAGCCATTGATTGCCTCCTTTGCAGGCAGTTGGTGGTTAGGCGCTGGCGGGGTCACTACCCCCGCCCTGCGCCGCTCTGAGTTCTTCCAGGCACTCGCGCAGCGCCTCGTATTCGTCGTCGGTCAGCCAGAACTTGCGCTGACGCATCCCTTTGGCCTTCATGCGCTCGGCATATCGCGCTTGGCGTACAGCGTCAGGGCTGGCCAGCAGGTCGCCGGTCTGCTTGTCCTTTTGGTCTTTCATATCAGTCACCGAGAAACTTGCCGGGGCCGAAAGGCTTGTCTTCGCGGAACAAGGACAAGGTGCAGCCGTCAAGGTTAAGGCTGGCCATATCAGCGACTGAAAAGCGGCCAGCCTCGGCCTTGTTTGACGTATAACCAAAGCCACCGGGCAACCAATAGCCCTTGTCACTTTTGATCAGGTAGACCGACCCTGAAGCGCTTTTTTCGTATTCCATAAAACCCCCTTCTGTGCGTGAGTACTCATAGTATAACAGGCGTGAGTACCAACGCAAGCACTACATGCTGAAATCTGGCCCGCCCCTGTCCATGCCTCGCTGGCGGGGTGCCGGTGCCCGTGCCAGCTCGGCCCGCGCAAGCTGGACGCTGGGCAGACCCATGACCTTGCTGACGGTGCGCTCGATGTAATCCGCTTCGTGGCCGGGCTTGCGCTCTGCCAGCGCTGGGCTGGCCTCGGCCATGGCCTTGCCGATTTCCTCGGCACTGCGGCCCCGGCTGGCCAGCTTCTGCGCGGCGATAAAGTCGCACTTGCTGAGGTCATCACCGAAGCGCTTGACCAGCCCGGCCATCTCGCTGCGGTACTCGTCCAGCGCCGTGCGCCGGTGGCGGCTAAGCTGCCGCTCGGGCAGTTCGAGGCTGGCCAGCCTGCGGGCCTTCTCCTGCTGCCGCTGGGCCTGCTCGATCTGCTGGCCAGCCTGCTGCACCAGCGCCGGGCCAGCGGTGGCGGTCTTGCCCTTGGATTCACGCAGCAGCACCCACGGCTGATAACCGGCGCGGGTGGTGTGCTTGTCCTTGCGGTTGGTGAAGCCCGCCAAGCGGCCATAGTGGCGGCTGTCGGCGCTGGCCGGGTCGGCGTCGTACTCGCTGGCCAGCGTCCGGGCAATCTGCCCCCGAAGTTCACCGCCTGCGGCGTCGGCCACCTTGACCCATGCCTGATAGTTCTTCGGGCTGGTTTCCACTACCAGGGCAGGCTCCCGGCCCTCGGCTTTCATGTCATCCAGGTCAAACTCGCTGAGGTCGTCCACCAGCACCAGACCATGCCGCTCCTGCTCGGCGGGCCTGATATACACGTCATTGCCCTGGGCATTCATCCGCTTGAGCCATGGCGTGTTCTGGAGCACTTCGGCGGCTGACCATTCCCGGTTCATCATCTGGCCGGTGGTGGCGTCCCTGACGCCGATATCGAAGCGCTCACAGCCCATGGCCTTGAGCTGTCGGCCTATGGCCTGCAAAGTCCTGTCGTTCTTCATCGGGCCACCAAGCGCAGCCAGATCGAGCCGTCCTCGGTTGTCAGTGGCGTCAGGTCGAGCAAGAGCAACGATGCGATCAGCAGCACCACCGTAGGCATCATGGAAGCCAGCATCACGGTTAGCCATAGCTTCCAGTGCCACCCCCGCGACGCGCTCCGGGCGCTCTGCGCGGCGCTGCTCACCTCGGCGGCTACCTCCCGCAACTCTTTGGCCAGCTCCACCCATGCCGCCCCTGTCTGGCGCTGGGCTTTCAGCCACTCCGCCGCCTGCGCCTCGCTGGCCTGCTGGGTCTGGCTCATGACCTGCCGGGCTTCGTCGGCCAGTGTCGCCATGCTCTGGGCCAGCGGTTCGATCTGCTCCGCTAACTCGTTGATGCCTCTGGATTTCTTCACTCTGTCGATTGCGTTCATGGTCTATTGCCTCCCGGTATTCCTGTAAGTCGATGATCTGGGCGTTGGCGGTGTCGATGTTCAGGGCCACGTCTGCCCGGTCGGTGCGGATGCCCCGGCCTTCCATCTCCACCACGTTCGGCCCCAGGTGAACACCGGGCAGGCGCTCGATGCCCTGCGCCTCAAGTGTTCTGTGGTCAATGCGGGCGTCGTGGCCAGCCCGCTCTAATGCCCGGTTGGCATGGTCGGCCCATGCCTCGCGGGTCTGCTCAAGCCATGCCTTGGGCTTGAGCGCTTCGGTCTTCTGTGCCCCGCCCTTCTCCGGGGTCTTGCCGTTGTACCGCTTGAACCACTGAGCGGCGGGCCGCTCGATGCCGTCATTGATCCGCTCGGAGATCATCAGGTGGCAGTGCGGGTTCTCGCCGCCACCGGCATGGATGGCCAGCGTATACGGCAGGCGCTCGGCACCGGTCAGGTGCTGGGCGAACTCGGACGCCAGCGCCTTCTGCTGGTCGAGGGTCAGCTCGACCGGCAGGGCAAATTCGACCTCCTTGAACAGCCGCCCATTGGCGCGTTCATACAGGTCGGCAGCATCCCAGTAGTCGGCGGGCCGCTCGACGAACTCCGGCATGTGCCCGGATTCGGCGTGCAAGACTTCATCCATGTCGCGGGCATACTTGCCTTCGCGCTGGATGTAGTCGGCCTTGGCCCTGGCCGATTGGCCGCCCGACCTGCTGCCGGTTTTCGCCGTAAGGTGATAAATCGCCATGCTGCCTCGCTGTTGCTTTTGCTTTTCGGCTCCATGCAATGGCCCTCGGAGAGCGCACCGCCCGAAGGGTGGCCGTTAGGCCAGTTTCTCGAAGAGAAACCGGTAAGTGCGCCCTCCCCTACAAAGTAGGGTCGGGATTGCCGCCGCTGTGCCTCCATGATAGCCTACGAGACAGCACATTAACAATGGGGTGTCAAGATGGTTAAGGGGAGCAACAAGGCGGCGGATCGGCTGGCCAAGCTCGAAGAACAACGAGCGCGAATCAATGCCGAAATTCAGCGGGTGCGGGCAAGGGAACAGCAGCAAGAGCGCAAGAACGAAACAAGGCGCAAGGTGCTGGTGGGGGCCATGATTTTGGCCAAGGTGAACAGCAGCGAGTGGCCGGAGGATCGGCTCATGGCGGCAATGGATGCGTACCTTGAACGCGACCACGACCGCGCCTTGTTCGGTCTGCCGCCACGCCAGAAGGATGAGCCGGGCTGAATGATCGACCGAGACAGGCCCTGCGGGGCTGCACACGCGCCCCCACCCTTCGGGTAGGGGGAAAGGCCGCTAAAGCGGCTAAAAGCGCTCCAGCGTATTTCTGCGGGGTTTGGTGTGGGGTTTAGCGGGCTTTGCCCGCCTTTCCCCCTGCCGCGCAGCGGTGGGGCGGTGTGTAGCCTAGCGCAGCGAATAGACCAGCTATCCGGCCTCTGGCCGGGCATATTGGGCAAGGGCAGCAGCGCCCCACAAGGGCGCTGATAACCGCGCCTAGTGGATTATTCTTAGATAATCATGGATGGATTTTTCCAACACCCCGCCAGCCCCCGCCCCTGCTGGGTTTGCAGGTTTGGGGGCGTGACAGTTATTGCAGGGGTTCGTGACAGTTATTGCAGGGGGGCGTGACAGTTATTGCAGGGGTTCGTGACAGTTAGTACGGGAGTGACGGGCACTGGCTGGCAATGTCTAGCAACGGCAGGCATTTCGGCTGAGGGTAAAAGAACTTTCCGCTAAGCGATAGACTGTATGTAAACACAGTATTGCAAGGACGCGGAACATGCCTCATGTGGCGGCCAGGACGGCCAGCCGGGATCGGGATACTGGTCGTTACCAGAGCCACCGACCCGAGCAAACCCTTCTCTATCAGATCGTTGACGAGTATTACCCGGCATTCGCTGCGCTTATGGCAGAGCAGGGAAAGGAATTGCCGGGCTATGTGCAACGGGAATTTGAAGAATTTCTCCAATGCGGGCGGCTGGAGCATGGCTTTCTACGGGTTCGCTGCGAGTCTTGCCACGCCGAGCACCTGGTCGCTTTCAGCTGTAATCCGGGCAGCGCAACGGAACATTCATCAGTGTAAAAATGGAATCAATAAAGCCCTGCGCAGCGCGCAGGGTCAGCCTGAATACGCGTTTAATGACCAGCACAGTCGTGATGGCAAGGTCAGAATAGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTTGGTGATTTTGAACTTTTGCTTTGCCACGGAACGGTCTGCGTTGTCGGGAAGATGCGTGATCTGATCCTTCAACTCAGCAAAAGTTCGATTTATTCAACAAAGCCGCCGTCCCGTCAAGTCAGCGTAATGCTCTGCCAGTGTTACAACCAATTAACCAATTCTGATTAGAAAAACTCATCGAGCATCAAATGAAACTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGCTTATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCGGAAGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTACCTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGTCGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGAGCAAGACGTTTCCCGTTGAATATGGCTCATAGCACCCCTTGTATTACTGTTTATGTAAGCAGA
Plasmid Elements
primer_bind 1079..1095
/note="常见的测序引物，多种相似变体之一"
RBS 1314..1336
/note="噬菌体T7基因10的高效核糖体结合位点（Olins和Rangwala，1989）"
GFPuv 1344..2060
/codon_start=1
/product="GFP variant optimized for excitation by UV light"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(2109..2125)
/note="常见的测序引物，多种相似变体之一"
lac operator 2133..2149
/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(2157..2187)
/note="大肠杆菌lac操纵子的启动子"
CAP binding site 2202..2223
/bound_moiety="E. coli catabolite activator protein"
/note="CAP结合在cAMP存在下激活转录。"
CDS complement(3725..4564)
/codon_start=1
/gene="repA"
/product="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS complement(5075..6046)
/codon_start=1
/gene="repB"
/product="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
oriT 7285..7372
/note="宽宿主范围质粒RSF1010的转移起源（Scholz等人，1989）"
RSF1010 oriV 7714..8108
/note="宽宿主范围质粒RSF1010的复制起点；需要RSF1010 RepA/B/C蛋白进行复制（Scholz等人，1989）"
KanR complement(8922..9737)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="赋予细菌对卡那霉素的抗性或真核生物对G418（遗传霉素（R））的抗性"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
