| 质粒名称: | pBTK800 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 179209 |
| 存储实验室: | Jeffrey Barrick |
|
载体骨架基本信息 |
|
| 载体名称: | RSF1010 |
| 完整质粒大小: | 8415 bp |
| 载体抗性: | Spectinomycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | GFP dropout |
| 物种来源: | Synthetic |
| 插入大小: | 1012 bp |
|
文献引用方法 |
|
| 材料和方法部分: | pBTK800 was a gift from Jeffrey Barrick (Addgene plasmid # 179209 ; http://n2t.net/addgene:179209 ; RRID:Addgene_179209) |
| 参考文献部分: | Honey bee functional genomics using symbiont-mediated RNAi. Lariviere PJ, Leonard SP, Horak RD, Powell JE, Barrick JE. Nat Protoc. 2023 Mar;18(3):902-928. doi: 10.1038/s41596-022-00778-4. Epub 2022 Dec 2. 10.1038/s41596-022-00778-4 PubMed 36460809 |
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 pBTK800 8415 bp ds-DNA circular SYN 10-MAY-2022
DEFINITION GFP dropout for assembling a double-stranded RNA expression plasmid.
ACCESSION .
VERSION .
KEYWORDS pBTK800
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8415)
TITLE Symbiont-mediated RNA interference for honey bee functional genomics
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8415)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 10, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8415
/organism="synthetic DNA construct"
/mol_type="other DNA"
oriT 2091..2178
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
rep_origin 2520..2914
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
RBS 3831..3842
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3849..4562
/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"
terminator 4574..4645
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4661..4688
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(4937..5728)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
CDS complement(6946..7785)
/codon_start=1
/gene="repA"
/product="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepA
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS complement(8296..852)
/codon_start=1
/gene="repB"
/product="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepB
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
ORIGIN
1 ctcgatgtaa tccgcttcgt ggccgggctt gcgctctgcc agcgctgggc tggcctcggc
61 catggccttg ccgatttcct cggcactgcg gccccggctg gccagcttct gcgcggcgat
121 aaagtcgcac ttgctgaggt catcaccgaa gcgcttgacc agcccggcca tctcgctgcg
181 gtactcgtcc agcgccgtgc gccggtggcg gctaagctgc cgctcgggca gttcgaggct
241 ggccagcctg cgggccttct cctgctgccg ctgggcctgc tcgatctgct ggccagcctg
301 ctgcaccagc gccgggccag cggtggcggt cttgcccttg gattcacgca gcagcaccca
361 cggctgataa ccggcgcggg tggtgtgctt gtccttgcgg ttggtgaagc ccgccaagcg
421 gccatagtgg cggctgtcgg cgctggccgg gtcggcgtcg tactcgctgg ccagcgtccg
481 ggcaatctgc ccccgaagtt caccgcctgc ggcgtcggcc accttgaccc atgcctgata
541 gttcttcggg ctggtttcca ctaccagggc aggctcccgg ccctcggctt tcatgtcatc
601 caggtcaaac tcgctgaggt cgtccaccag caccagacca tgccgctcct gctcggcggg
661 cctgatatac acgtcattgc cctgggcatt catccgcttg agccatggcg tgttctggag
721 cacttcggcg gctgaccatt cccggttcat catctggccg gtggtggcgt ccctgacgcc
781 gatatcgaag cgctcacagc ccatggcctt gagctgtcgg cctatggcct gcaaagtcct
841 gtcgttcttc atcgggccac caagcgcagc cagatcgagc cgtcctcggt tgtcagtggc
901 gtcaggtcga gcaagagcaa cgatgcgatc agcagcacca ccgtaggcat catggaagcc
961 agcatcacgg ttagccatag cttccagtgc cacccccgcg acgcgctccg ggcgctctgc
1021 gcggcgctgc tcacctcggc ggctacctcc cgcaactctt tggccagctc cacccatgcc
1081 gcccctgtct ggcgctgggc tttcagccac tccgccgcct gcgcctcgct ggcctgctgg
1141 gtctggctca tgacctgccg ggcttcgtcg gccagtgtcg ccatgctctg ggccagcggt
1201 tcgatctgct ccgctaactc gttgatgcct ctggatttct tcactctgtc gattgcgttc
1261 atggtctatt gcctcccggt attcctgtaa gtcgatgatc tgggcgttgg cggtgtcgat
1321 gttcagggcc acgtctgccc ggtcggtgcg gatgccccgg ccttccatct ccaccacgtt
1381 cggccccagg tgaacaccgg gcaggcgctc gatgccctgc gcctcaagtg ttctgtggtc
1441 aatgcgggcg tcgtggccag cccgctctaa tgcccggttg gcatggtcgg cccatgcctc
1501 gcgggtctgc tcaagccatg ccttgggctt gagcgcttcg gtcttctgtg ccccgccctt
1561 ctccggggtc ttgccgttgt accgcttgaa ccactgagcg gcgggccgct cgatgccgtc
1621 attgatccgc tcggagatca tcaggtggca gtgcgggttc tcgccgccac cggcatggat
1681 ggccagcgta tacggcaggc gctcggcacc ggtcaggtgc tgggcgaact cggacgccag
1741 cgccttctgc tggtcgaggg tcagctcgac cggcagggca aattcgacct ccttgaacag
1801 ccgcccattg gcgcgttcat acaggtcggc agcatcccag tagtcggcgg gccgctcgac
1861 gaactccggc atgtgcccgg attcggcgtg caagacttca tccatgtcgc gggcatactt
1921 gccttcgcgc tggatgtagt cggccttggc cctggccgat tggccgcccg acctgctgcc
1981 ggttttcgcc gtaaggtgat aaatcgccat gctgcctcgc tgttgctttt gcttttcggc
2041 tccatgcaat ggccctcgga gagcgcaccg cccgaagggt ggccgttagg ccagtttctc
2101 gaagagaaac cggtaagtgc gccctcccct acaaagtagg gtcgggattg ccgccgctgt
2161 gcctccatga tagcctacga gacagcacat taacaatggg gtgtcaagat ggttaagggg
2221 agcaacaagg cggcggatcg gctggccaag ctcgaagaac aacgagcgcg aatcaatgcc
2281 gaaattcagc gggtgcgggc aagggaacag cagcaagagc gcaagaacga aacaaggcgc
2341 aaggtgctgg tgggggccat gattttggcc aaggtgaaca gcagcgagtg gccggaggat
2401 cggctcatgg cggcaatgga tgcgtacctt gaacgcgacc acgaccgcgc cttgttcggt
2461 ctgccgccac gccagaagga tgagccgggc tgaatgatcg accgagacag gccctgcggg
2521 gctgcacacg cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg gctaaaagcg
2581 ctccagcgta tttctgcggg gtttggtgtg gggtttagcg ggctttgccc gcctttcccc
2641 ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca gctatccggc
2701 ctctggccgg gcatattggg caagggcagc agcgccccac aagggcgctg ataaccgcgc
2761 ctagtggatt attcttagat aatcatggat ggatttttcc aacaccccgc cagcccccgc
2821 ccctgctggg tttgcaggtt tgggggcgtg acagttattg caggggttcg tgacagttat
2881 tgcagggggg cgtgacagtt attgcagggg ttcgtgacag ttagtacggg agtgacgggc
2941 actggctggc aatgtctagc aacggcaggc atttcggctg agggtaaaag aactttccgc
3001 taagcgatag actgtatgta aacacagtat tgcaaggacg cggaacatgc ctcatgtggc
3061 ggccaggacg gccagccggg atcgggatac tggtcgttac cagagccacc gacccgagca
3121 aacccttctc tatcagatcg ttgacgagta ttacccggca ttcgctgcgc ttatggcaga
3181 gcagggaaag gaattgccgg gctatgtgca acgggaattt gaagaatttc tccaatgcgg
3241 gcggctggag catggctttc tacgggttcg ctgcgagtct tgccacgccg agcacctggt
3301 cgctttcaga aatcaatcta aagtatatat gagtaaactt ggtctgacag gcccctgaat
3361 tcgcatctag actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc
3421 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
3481 cccacagtac agacactgcg acaacgtggc aattcgtcgc aatacaacgg acagtcattc
3541 atctttctgc ccctccaaaa gcaaaaaccc gccgaagcgg gtttttacgt aaatcaggtg
3601 aaactgaccg actttggcag tttattcttg acatgtagtg agggggctgg tataatcaca
3661 tagtactgtt ttatgtgaga ccgaaagtga aacgtgattt catgcgtcat tttgaacatt
3721 ttgtaaatct tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt
3781 aacatcgcgg caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga
3841 aatactagat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac
3901 tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa
3961 ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc
4021 cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata
4081 tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga
4141 tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata
4201 ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg
4261 gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa
4321 aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc
4381 tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca
4441 atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata
4501 tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca
4561 aatgaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
4621 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
4681 tttctgcgtt tataggtckc anatmmaaca gtactatgtg attataccag ccccctcact
4741 acatgtcaag aataaactgc caaaggtaat cgttaatccg caaataacgt aaaaacccgc
4801 ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcagc tgcgtctcaa
4861 gcagttacag agatgttacg aacccccagg acatccgaga atgcgaggcg atggagggta
4921 caacccgagc ggccgcttat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
4981 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
5041 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
5101 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
5161 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
5221 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
5281 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
5341 caatgtcgat cgtggctggc tcgaagatac ccgcaagaat gtcattgcgc tgccattctc
5401 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
5461 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
5521 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
5581 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
5641 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
5701 atacttcggc gatcaccgct tccctcataa tgtttaactt tgttttaggg cgactgccct
5761 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
5821 gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga
5881 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
5941 agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc
6001 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggtagc agcgaagtcg
6061 aggcatttct gtcctggctg gtcatgacca aaatccctta acgtgagtca gcctgccgcc
6121 ttgggccggg tgatgtcgta cttgcccgcc gcgaactcgg ttaccgtcca gcccagcgcg
6181 accagctccg gcaacgcctc gcgcacccgc tggcggcgct tgcgcatggt cgaaccactg
6241 gcctctgacg gccagacata gccgcacaag gtatctatgg aagccttgcc ggttttgccg
6301 gggtcgatcc agccacacag ccgctggtgc agcaggcggg cggtttcgct gtccagcgcc
6361 cgcacctcgt ccatgctgat gcgcacatgc tggccgccac ccatgacggc ctgcgcgatc
6421 aaggggttca gggccacgta caggcgcccg tccgcctcgt cgctggcgta ctccgacagc
6481 agccgaaacc cctgccgctt gcggccattc tgggcgatga tggatacctt ccaaaggcgc
6541 tcgatgcagt cctgtatgtg cttgagcgcc ccaccactat cgacctctgc cccgatttcc
6601 tttgccagcg cccgatagct acctttgacc accatggcat cagcggtgac ggcctcccac
6661 ttgggttcca ggaacagccg gagctgccgt ccgccttcgg tcttgggttc cgggccaagc
6721 actaggccat taggcccagc catggccacc agcccttgca ggatgcgcag atcatcagcg
6781 cccagcggct ccgggccgct gaactcgatc cgcttgccgt cgccgtagtc atacgtcacg
6841 tccagcttgc tgcgcttgcg ctcgccccgc ttgagggcac ggaacaggcc gggggccaga
6901 cagtgcgccg ggtcgtgccg gacgtggctg aggctgtgct tgttcttagg cttcaccacg
6961 gggcaccccc ttgctcttgc gctgcctctc cagcacggcg ggcttgagca ccccgccgtc
7021 atgccgcctg aaccaccgat cagcgaacgg tgcgccatag ttggccttgc tcacaccgaa
7081 gcggacgaag aaccggcgct ggtcgtcgtc cacaccccat tcctcggcct cggcgctggt
7141 catgctcgac aggtaggact gccagcggat gttatcgacc agtaccgagc tgccccggct
7201 ggcctgctgc tggtcgcctg cgcccatcat ggccgcgccc ttgctggcat ggtgcaggaa
7261 cacgatagag cacccggtat cggcggcgat ggcctccatg cgaccgatga cctgggccat
7321 ggggccgctg gcgttttctt cctcgatgtg gaaccggcgc agcgtgtcca gcaccatcag
7381 gcggcggccc tcggcggcgc gcttgaggcc gtcgaaccac tccggggcca tgatgttggg
7441 caggctgccg atcagcggct ggatcagcag gccgtcagcc acggcttgcc gttcctcggc
7501 gctgaggtgc gccccaaggg cgtgcaggcg gtgatgaatg gcggtgggcg ggtcttcggc
7561 gggcaggtag atcaccgggc cggtgggcag ttcgcccacc tccagcagat ccggcccgcc
7621 tgcaatctgt gcggccagtt gcagggccag catggattta ccggcaccac cgggcgacac
7681 cagcgccccg accgtaccgg ccaccatgtt gggcaaaacg tagtccagcg gtggcggcgc
7741 tgctgcgaac gcctccagaa tattgatagg cttatgggta gccattgatt gcctcctttg
7801 caggcagttg gtggttaggc gctggcgggg tcactacccc cgccctgcgc cgctctgagt
7861 tcttccaggc actcgcgcag cgcctcgtat tcgtcgtcgg tcagccagaa cttgcgctga
7921 cgcatccctt tggccttcat gcgctcggca tatcgcgctt ggcgtacagc gtcagggctg
7981 gccagcaggt cgccggtctg cttgtccttt tggtctttca tatcagtcac cgagaaactt
8041 gccggggccg aaaggcttgt cttcgcggaa caaggacaag gtgcagccgt caaggttaag
8101 gctggccata tcagcgactg aaaagcggcc agcctcggcc ttgtttgacg tataaccaaa
8161 gccaccgggc aaccaatagc ccttgtcact tttgatcagg tagaccgacc ctgaagcgct
8221 tttttcgtat tccataaaac ccccttctgt gcgtgagtac tcatagtata acaggcgtga
8281 gtaccaacgc aagcactaca tgctgaaatc tggcccgccc ctgtccatgc ctcgctggcg
8341 gggtgccggt gcccgtgcca gctcggcccg cgcaagctgg acgctgggca gacccatgac
8401 cttgctgacg gtgcg
//
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