| 质粒名称: | pBTK520 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 110604 |
| 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: | 8272 bp |
| 载体抗性: | Spectinomycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | Synthetic |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pBTK520 was a gift from Jeffrey Barrick (Addgene plasmid # 110604 ; http://n2t.net/addgene:110604 ; RRID:Addgene_110604) |
| 参考文献部分: | Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar S, Davies BW, Ellington AD, Moran NA, Barrick JE. ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 10.1021/acssynbio.7b00399. Epub 2018 Apr 13. 10.1021/acssynbio.7b00399 PubMed 29608282 |
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 pBTK520 8272 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION BTK assembled plasmid- broad-host-range GFP expression from PA1
promoter (spectinomycin resistance).
ACCESSION .
VERSION .
KEYWORDS pBTK520
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8272)
AUTHORS Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar
S, Davies BW, Ellington AD, Moran NA, Barrick JE
TITLE Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit
for Modular Assembly of Broad-Host-Range Plasmids.
JOURNAL ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi:
10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
PUBMED 29608282
REFERENCE 2 (bases 1 to 8272)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8272
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 913..1752
/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 2970..3761
/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"
terminator 3780..3807
/label=T7Te terminator
/note="phage T7 early transcription terminator"
terminator 3823..3894
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 4910..4934
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
rep_origin 5641..6035
/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)"
oriT 6377..6464
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 7703..402
/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 gagcaggccc agcggcagca ggagaaggcc cgcaggctgg ccagcctcga actgcccgag
61 cggcagctta gccgccaccg gcgcacggcg ctggacgagt accgcagcga gatggccggg
121 ctggtcaagc gcttcggtga tgacctcagc aagtgcgact ttatcgccgc gcagaagctg
181 gccagccggg gccgcagtgc cgaggaaatc ggcaaggcca tggccgaggc cagcccagcg
241 ctggcagagc gcaagcccgg ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc
301 atgggtctgc ccagcgtcca gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc
361 cagcgaggca tggacagggg cgggccagat ttcagcatgt agtgcttgcg ttggtactca
421 cgcctgttat actatgagta ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc
481 gcttcagggt cggtctacct gatcaaaagt gacaagggct attggttgcc cggtggcttt
541 ggttatacgt caaacaaggc cgaggctggc cgcttttcag tcgctgatat ggccagcctt
601 aaccttgacg gctgcacctt gtccttgttc cgcgaagaca agcctttcgg ccccggcaag
661 tttctcggtg actgatatga aagaccaaaa ggacaagcag accggcgacc tgctggccag
721 ccctgacgct gtacgccaag cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca
781 gcgcaagttc tggctgaccg acgacgaata cgaggcgctg cgcgagtgcc tggaagaact
841 cagagcggcg cagggcgggg gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa
901 aggaggcaat caatggctac ccataagcct atcaatattc tggaggcgtt cgcagcagcg
961 ccgccaccgc tggactacgt tttgcccaac atggtggccg gtacggtcgg ggcgctggtg
1021 tcgcccggtg gtgccggtaa atccatgctg gccctgcaac tggccgcaca gattgcaggc
1081 gggccggatc tgctggaggt gggcgaactg cccaccggcc cggtgatcta cctgcccgcc
1141 gaagacccgc ccaccgccat tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc
1201 gaggaacggc aagccgtggc tgacggcctg ctgatccagc cgctgatcgg cagcctgccc
1261 aacatcatgg ccccggagtg gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg
1321 atggtgctgg acacgctgcg ccggttccac atcgaggaag aaaacgccag cggccccatg
1381 gcccaggtca tcggtcgcat ggaggccatc gccgccgata ccgggtgctc tatcgtgttc
1441 ctgcaccatg ccagcaaggg cgcggccatg atgggcgcag gcgaccagca gcaggccagc
1501 cggggcagct cggtactggt cgataacatc cgctggcagt cctacctgtc gagcatgacc
1561 agcgccgagg ccgaggaatg gggtgtggac gacgaccagc gccggttctt cgtccgcttc
1621 ggtgtgagca aggccaacta tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac
1681 ggcggggtgc tcaagcccgc cgtgctggag aggcagcgca agagcaaggg ggtgccccgt
1741 ggtgaagcct aagaacaagc acagcctcag ccacgtccgg cacgacccgg cgcactgtct
1801 ggcccccggc ctgttccgtg ccctcaagcg gggcgagcgc aagcgcagca agctggacgt
1861 gacgtatgac tacggcgacg gcaagcggat cgagttcagc ggcccggagc cgctgggcgc
1921 tgatgatctg cgcatcctgc aagggctggt ggccatggct gggcctaatg gcctagtgct
1981 tggcccggaa cccaagaccg aaggcggacg gcagctccgg ctgttcctgg aacccaagtg
2041 ggaggccgtc accgctgatg ccatggtggt caaaggtagc tatcgggcgc tggcaaagga
2101 aatcggggca gaggtcgata gtggtggggc gctcaagcac atacaggact gcatcgagcg
2161 cctttggaag gtatccatca tcgcccagaa tggccgcaag cggcaggggt ttcggctgct
2221 gtcggagtac gccagcgacg aggcggacgg gcgcctgtac gtggccctga accccttgat
2281 cgcgcaggcc gtcatgggtg gcggccagca tgtgcgcatc agcatggacg aggtgcgggc
2341 gctggacagc gaaaccgccc gcctgctgca ccagcggctg tgtggctgga tcgaccccgg
2401 caaaaccggc aaggcttcca tagatacctt gtgcggctat gtctggccgt cagaggccag
2461 tggttcgacc atgcgcaagc gccgccagcg ggtgcgcgag gcgttgccgg agctggtcgc
2521 gctgggctgg acggtaaccg agttcgcggc gggcaagtac gacatcaccc ggcccaaggc
2581 ggcaggctga ctcacgttaa gggattttgg tcatgaccag ccaggacaga aatgcctcga
2641 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
2701 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
2761 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
2821 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
2881 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
2941 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
3001 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
3061 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
3121 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
3181 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
3241 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
3301 aatggcagcg caatgacatt cttgcgggta tcttcgagcc agccacgatc gacattgatc
3361 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
3421 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
3481 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
3541 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
3601 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
3661 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
3721 actacgtgaa aggcgagatc accaaggtag tcggcaaata agcggccgct cggtataaac
3781 gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt tcaccgacaa
3841 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
3901 ggtgtactgc agtgcactag ttaactgcct tgatctgtcg ggtgtctgct tctctcacag
3961 gcttaggtgg atctgctgtc cccagtcagt gcggtcatca tgcagtcatc cgagcgtgta
4021 ttgcacacat ttgtttggcg catgatgagc cgtgatgacc ccgtcgtctc attggcagct
4081 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg
4141 ttatttgcgg attaacgatt acggatttat ttgtatagtt catccatgcc gtgtgtaatc
4201 ccagcagccg tgacaaactc aagaaggacc atgtggtcgc gcttttcgtt gggatctttc
4261 gaaagggcag attgagtgga caggtaatgg ttgtctggta acagcacagg cccatcgcca
4321 attggggtgt tttgttggta atggtcggct agttgaacgc ttccatcctc aatgttgtgt
4381 ctgatcttga agttaacctt gataccgttc ttttgcttgt cggccataat gtacacatta
4441 tgggagttat agttatattc cagtttgtgg cctaaaatat ttccatcttc cttaaagtca
4501 atacccttta actcgattct attaacaagg gtatcaccct cgaacttcac ttcggcgcgt
4561 gtcttatagt tgccgtcatc cttaaaaaag atggttctct cctgtacata gccttcgggc
4621 attgcactct tgaaaaagtc gtgctgtttc atgtgatctg ggtagcgcgc aaagcactga
4681 accccatacc cgaacgtagt gaccagtgtt ggccatggaa caggtagttt acccgtggtg
4741 caaataaatt taagggtcag cttgccatat gttgcgtcgc cttcaccctc tccactgaca
4801 gagaacttgt gcccattaac atcaccgtct aattccacga ggattgggac aactcctgtg
4861 aaaagctctt ctcctttact catatgtaat atctcctctg tttctgtatg gaattgttat
4921 ccgctcacaa ttcctcgccg tgtccctctc gatggctgta agtatcctat aggttagact
4981 ttaagtcaat actctttttg ataacgttcg tctgcaatta tcggccagtt cagtgagacg
5041 gtattgcgac gaattgccac gttgtcgcag tgtctgtact gtgggttaat gtctgtgatc
5101 gccgtcgtga gttcgtgtca tccgcgcatg attcagatgg tcctggagat cgttgcttgt
5161 accggctgtt tgtggctcta ccatctagat gcgaattcag gggcctgtca gaccaagttt
5221 actcatatat actttagatt gatttctgaa agcgaccagg tgctcggcgt ggcaagactc
5281 gcagcgaacc cgtagaaagc catgctccag ccgcccgcat tggagaaatt cttcaaattc
5341 ccgttgcaca tagcccggca attcctttcc ctgctctgcc ataagcgcag cgaatgccgg
5401 gtaatactcg tcaacgatct gatagagaag ggtttgctcg ggtcggtggc tctggtaacg
5461 accagtatcc cgatcccggc tggccgtcct ggccgccaca tgaggcatgt tccgcgtcct
5521 tgcaatactg tgtttacata cagtctatcg cttagcggaa agttctttta ccctcagccg
5581 aaatgcctgc cgttgctaga cattgccagc cagtgcccgt cactcccgta ctaactgtca
5641 cgaacccctg caataactgt cacgcccccc tgcaataact gtcacgaacc cctgcaataa
5701 ctgtcacgcc cccaaacctg caaacccagc aggggcgggg gctggcgggg tgttggaaaa
5761 atccatccat gattatctaa gaataatcca ctaggcgcgg ttatcagcgc ccttgtgggg
5821 cgctgctgcc cttgcccaat atgcccggcc agaggccgga tagctggtct attcgctgcg
5881 ctaggctaca caccgcccca ccgctgcgcg gcagggggaa aggcgggcaa agcccgctaa
5941 accccacacc aaaccccgca gaaatacgct ggagcgcttt tagccgcttt agcggccttt
6001 ccccctaccc gaagggtggg ggcgcgtgtg cagccccgca gggcctgtct cggtcgatca
6061 ttcagcccgg ctcatccttc tggcgtggcg gcagaccgaa caaggcgcgg tcgtggtcgc
6121 gttcaaggta cgcatccatt gccgccatga gccgatcctc cggccactcg ctgctgttca
6181 ccttggccaa aatcatggcc cccaccagca ccttgcgcct tgtttcgttc ttgcgctctt
6241 gctgctgttc ccttgcccgc acccgctgaa tttcggcatt gattcgcgct cgttgttctt
6301 cgagcttggc cagccgatcc gccgccttgt tgctcccctt aaccatcttg acaccccatt
6361 gttaatgtgc tgtctcgtag gctatcatgg aggcacagcg gcggcaatcc cgaccctact
6421 ttgtagggga gggcgcactt accggtttct cttcgagaaa ctggcctaac ggccaccctt
6481 cgggcggtgc gctctccgag ggccattgca tggagccgaa aagcaaaagc aacagcgagg
6541 cagcatggcg atttatcacc ttacggcgaa aaccggcagc aggtcgggcg gccaatcggc
6601 cagggccaag gccgactaca tccagcgcga aggcaagtat gcccgcgaca tggatgaagt
6661 cttgcacgcc gaatccgggc acatgccgga gttcgtcgag cggcccgccg actactggga
6721 tgctgccgac ctgtatgaac gcgccaatgg gcggctgttc aaggaggtcg aatttgccct
6781 gccggtcgag ctgaccctcg accagcagaa ggcgctggcg tccgagttcg cccagcacct
6841 gaccggtgcc gagcgcctgc cgtatacgct ggccatccat gccggtggcg gcgagaaccc
6901 gcactgccac ctgatgatct ccgagcggat caatgacggc atcgagcggc ccgccgctca
6961 gtggttcaag cggtacaacg gcaagacccc ggagaagggc ggggcacaga agaccgaagc
7021 gctcaagccc aaggcatggc ttgagcagac ccgcgaggca tgggccgacc atgccaaccg
7081 ggcattagag cgggctggcc acgacgcccg cattgaccac agaacacttg aggcgcaggg
7141 catcgagcgc ctgcccggtg ttcacctggg gccgaacgtg gtggagatgg aaggccgggg
7201 catccgcacc gaccgggcag acgtggccct gaacatcgac accgccaacg cccagatcat
7261 cgacttacag gaataccggg aggcaataga ccatgaacgc aatcgacaga gtgaagaaat
7321 ccagaggcat caacgagtta gcggagcaga tcgaaccgct ggcccagagc atggcgacac
7381 tggccgacga agcccggcag gtcatgagcc agacccagca ggccagcgag gcgcaggcgg
7441 cggagtggct gaaagcccag cgccagacag gggcggcatg ggtggagctg gccaaagagt
7501 tgcgggaggt agccgccgag gtgagcagcg ccgcgcagag cgcccggagc gcgtcgcggg
7561 ggtggcactg gaagctatgg ctaaccgtga tgctggcttc catgatgcct acggtggtgc
7621 tgctgatcgc atcgttgctc ttgctcgacc tgacgccact gacaaccgag gacggctcga
7681 tctggctgcg cttggtggcc cgatgaagaa cgacaggact ttgcaggcca taggccgaca
7741 gctcaaggcc atgggctgtg agcgcttcga tatcggcgtc agggacgcca ccaccggcca
7801 gatgatgaac cgggaatggt cagccgccga agtgctccag aacacgccat ggctcaagcg
7861 gatgaatgcc cagggcaatg acgtgtatat caggcccgcc gagcaggagc ggcatggtct
7921 ggtgctggtg gacgacctca gcgagtttga cctggatgac atgaaagccg agggccggga
7981 gcctgccctg gtagtggaaa ccagcccgaa gaactatcag gcatgggtca aggtggccga
8041 cgccgcaggc ggtgaacttc gggggcagat tgcccggacg ctggccagcg agtacgacgc
8101 cgacccggcc agcgccgaca gccgccacta tggccgcttg gcgggcttca ccaaccgcaa
8161 ggacaagcac accacccgcg ccggttatca gccgtgggtg ctgctgcgtg aatccaaggg
8221 caagaccgcc accgctggcc cggcgctggt gcagcaggct ggccagcaga tc
//
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