| 质粒名称: | pBTK570 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110615 |
| 存储实验室: | Jeffrey Barrick |
|
载体骨架基本信息 |
|
| 载体名称: | RSF1010 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 大观霉素 |
| 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: | E2-Crimson |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pBTK570 was a gift from Jeffrey Barrick (Addgene plasmid # 110615 ; http://n2t.net/addgene:110615 ; RRID:Addgene_110615) |
| 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 pBTK570 7999 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION BTK assembled broad-host-range plasmid encoding E2-Crimson protein
driven by PA3 promoter.
ACCESSION .
VERSION .
KEYWORDS pBTK570
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7999)
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 7999)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7999
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 617..1291
/codon_start=1
/product="far-red noncytotoxic tetrameric variant of DsRed
fluorescent protein (Strack et al., 2009)"
/label=E2-Crimson
/note="mammalian codon-optimized"
/translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
LFQ"
primer_bind 1015..1034
/label=mCherry-F
/note="mCherry, forward primer"
CDS complement(1491..2282)
/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(3500..4339)
/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(4850..5821)
/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"
oriT 7060..7147
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
rep_origin 7489..7883
/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)"
ORIGIN
1 tgcaaggacg cggaacatgc ctcatgtggc ggccaggacg gccagccggg atcgggatac
61 tggtcgttac cagagccacc gacccgagca aacccttctc tatcagatcg ttgacgagta
121 ttacccggca ttcgctgcgc ttatggcaga gcagggaaag gaattgccgg gctatgtgca
181 acgggaattt gaagaatttc tccaatgcgg gcggctggag catggctttc tacgggttcg
241 ctgcgagtct tgccacgccg agcacctggt cgctttcaga aatcaatcta aagtatatat
301 gagtaaactt ggtctgacag gcccctgaat tcgcatctag atggtagagc cacaaacagc
361 cggtacaagc aacgatctcc aggaccatct gaatcatgcg cggatgacac gaactcacga
421 cggcgatcac agacattaac ccacagtaca gacactgcga caacgtggca attcgtcgca
481 ataccgtctc actgaactgg ccgataattg cagacgaacg ggtgaaacaa aacggttgac
541 aacatgaagt aaacacggta cgatgtacca catgaaacga cagtgagtca atacagaaac
601 agaggagata ttacatatgg atagcactga gaacgtcatc aagcccttca tgcgcttcaa
661 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg tgggcgaggg
721 caagccctac gagggcaccc agaccgccaa gctgcaagtg accaagggcg gccccctgcc
781 cttcgcctgg gacatcctgt ccccccagtt cttctacggc tccaaggcgt acatcaagca
841 ccccgccgac atccccgact acctcaagca gtccttcccc gagggcttca agtgggagcg
901 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
961 cggcaccctc atctaccacg tgaagttcat cggcgtgaac ttcccctccg acggccccgt
1021 aatgcagaag aagactctgg gctgggagcc ctccactgag cgcaactacc cccgcgacgg
1081 cgtgctgaag ggcgagaacc acatggcgct gaagctgaag ggcggcggcc actacctgtg
1141 tgagttcaag tccatctaca tggccaagaa gcccgtgaag ctgcccggct accactacgt
1201 ggactacaag ctcgacatca cctcccacaa cgaggactac accgtggtgg agcagtacga
1261 gcgcgccgag gcccgccacc acctgttcca gactcacggt atggacgaat tgtacaagca
1321 cgacgaattg taaatccgta atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg
1381 gtttttttat ggggggagtt tagggaaaga gcatttgtca gctggatcag cagatgggaa
1441 ccggatactc ctcagcctcc agagtagcca atgagacgcc gagcggccgc ttatttgccg
1501 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
1561 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
1621 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
1681 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
1741 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
1801 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
1861 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
1921 atacccgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
1981 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
2041 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
2101 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
2161 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
2221 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
2281 ataatgttta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
2341 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
2401 tgtaccccaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg
2461 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagc
2521 ttacgaaccg aacaggctta tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg
2581 cgtcacccgg caaccttggg tagcagcgaa gtcgaggcat ttctgtcctg gctggtcatg
2641 accaaaatcc cttaacgtga gtcagcctgc cgccttgggc cgggtgatgt cgtacttgcc
2701 cgccgcgaac tcggttaccg tccagcccag cgcgaccagc tccggcaacg cctcgcgcac
2761 ccgctggcgg cgcttgcgca tggtcgaacc actggcctct gacggccaga catagccgca
2821 caaggtatct atggaagcct tgccggtttt gccggggtcg atccagccac acagccgctg
2881 gtgcagcagg cgggcggttt cgctgtccag cgcccgcacc tcgtccatgc tgatgcgcac
2941 atgctggccg ccacccatga cggcctgcgc gatcaagggg ttcagggcca cgtacaggcg
3001 cccgtccgcc tcgtcgctgg cgtactccga cagcagccga aacccctgcc gcttgcggcc
3061 attctgggcg atgatggata ccttccaaag gcgctcgatg cagtcctgta tgtgcttgag
3121 cgccccacca ctatcgacct ctgccccgat ttcctttgcc agcgcccgat agctaccttt
3181 gaccaccatg gcatcagcgg tgacggcctc ccacttgggt tccaggaaca gccggagctg
3241 ccgtccgcct tcggtcttgg gttccgggcc aagcactagg ccattaggcc cagccatggc
3301 caccagccct tgcaggatgc gcagatcatc agcgcccagc ggctccgggc cgctgaactc
3361 gatccgcttg ccgtcgccgt agtcatacgt cacgtccagc ttgctgcgct tgcgctcgcc
3421 ccgcttgagg gcacggaaca ggccgggggc cagacagtgc gccgggtcgt gccggacgtg
3481 gctgaggctg tgcttgttct taggcttcac cacggggcac ccccttgctc ttgcgctgcc
3541 tctccagcac ggcgggcttg agcaccccgc cgtcatgccg cctgaaccac cgatcagcga
3601 acggtgcgcc atagttggcc ttgctcacac cgaagcggac gaagaaccgg cgctggtcgt
3661 cgtccacacc ccattcctcg gcctcggcgc tggtcatgct cgacaggtag gactgccagc
3721 ggatgttatc gaccagtacc gagctgcccc ggctggcctg ctgctggtcg cctgcgccca
3781 tcatggccgc gcccttgctg gcatggtgca ggaacacgat agagcacccg gtatcggcgg
3841 cgatggcctc catgcgaccg atgacctggg ccatggggcc gctggcgttt tcttcctcga
3901 tgtggaaccg gcgcagcgtg tccagcacca tcaggcggcg gccctcggcg gcgcgcttga
3961 ggccgtcgaa ccactccggg gccatgatgt tgggcaggct gccgatcagc ggctggatca
4021 gcaggccgtc agccacggct tgccgttcct cggcgctgag gtgcgcccca agggcgtgca
4081 ggcggtgatg aatggcggtg ggcgggtctt cggcgggcag gtagatcacc gggccggtgg
4141 gcagttcgcc cacctccagc agatccggcc cgcctgcaat ctgtgcggcc agttgcaggg
4201 ccagcatgga tttaccggca ccaccgggcg acaccagcgc cccgaccgta ccggccacca
4261 tgttgggcaa aacgtagtcc agcggtggcg gcgctgctgc gaacgcctcc agaatattga
4321 taggcttatg ggtagccatt gattgcctcc tttgcaggca gttggtggtt aggcgctggc
4381 ggggtcacta cccccgccct gcgccgctct gagttcttcc aggcactcgc gcagcgcctc
4441 gtattcgtcg tcggtcagcc agaacttgcg ctgacgcatc cctttggcct tcatgcgctc
4501 ggcatatcgc gcttggcgta cagcgtcagg gctggccagc aggtcgccgg tctgcttgtc
4561 cttttggtct ttcatatcag tcaccgagaa acttgccggg gccgaaaggc ttgtcttcgc
4621 ggaacaagga caaggtgcag ccgtcaaggt taaggctggc catatcagcg actgaaaagc
4681 ggccagcctc ggccttgttt gacgtataac caaagccacc gggcaaccaa tagcccttgt
4741 cacttttgat caggtagacc gaccctgaag cgcttttttc gtattccata aaaccccctt
4801 ctgtgcgtga gtactcatag tataacaggc gtgagtacca acgcaagcac tacatgctga
4861 aatctggccc gcccctgtcc atgcctcgct ggcggggtgc cggtgcccgt gccagctcgg
4921 cccgcgcaag ctggacgctg ggcagaccca tgaccttgct gacggtgcgc tcgatgtaat
4981 ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct ggcctcggcc atggccttgc
5041 cgatttcctc ggcactgcgg ccccggctgg ccagcttctg cgcggcgata aagtcgcact
5101 tgctgaggtc atcaccgaag cgcttgacca gcccggccat ctcgctgcgg tactcgtcca
5161 gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag ttcgaggctg gccagcctgc
5221 gggccttctc ctgctgccgc tgggcctgct cgatctgctg gccagcctgc tgcaccagcg
5281 ccgggccagc ggtggcggtc ttgcccttgg attcacgcag cagcacccac ggctgataac
5341 cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc cgccaagcgg ccatagtggc
5401 ggctgtcggc gctggccggg tcggcgtcgt actcgctggc cagcgtccgg gcaatctgcc
5461 cccgaagttc accgcctgcg gcgtcggcca ccttgaccca tgcctgatag ttcttcgggc
5521 tggtttccac taccagggca ggctcccggc cctcggcttt catgtcatcc aggtcaaact
5581 cgctgaggtc gtccaccagc accagaccat gccgctcctg ctcggcgggc ctgatataca
5641 cgtcattgcc ctgggcattc atccgcttga gccatggcgt gttctggagc acttcggcgg
5701 ctgaccattc ccggttcatc atctggccgg tggtggcgtc cctgacgccg atatcgaagc
5761 gctcacagcc catggccttg agctgtcggc ctatggcctg caaagtcctg tcgttcttca
5821 tcgggccacc aagcgcagcc agatcgagcc gtcctcggtt gtcagtggcg tcaggtcgag
5881 caagagcaac gatgcgatca gcagcaccac cgtaggcatc atggaagcca gcatcacggt
5941 tagccatagc ttccagtgcc acccccgcga cgcgctccgg gcgctctgcg cggcgctgct
6001 cacctcggcg gctacctccc gcaactcttt ggccagctcc acccatgccg cccctgtctg
6061 gcgctgggct ttcagccact ccgccgcctg cgcctcgctg gcctgctggg tctggctcat
6121 gacctgccgg gcttcgtcgg ccagtgtcgc catgctctgg gccagcggtt cgatctgctc
6181 cgctaactcg ttgatgcctc tggatttctt cactctgtcg attgcgttca tggtctattg
6241 cctcccggta ttcctgtaag tcgatgatct gggcgttggc ggtgtcgatg ttcagggcca
6301 cgtctgcccg gtcggtgcgg atgccccggc cttccatctc caccacgttc ggccccaggt
6361 gaacaccggg caggcgctcg atgccctgcg cctcaagtgt tctgtggtca atgcgggcgt
6421 cgtggccagc ccgctctaat gcccggttgg catggtcggc ccatgcctcg cgggtctgct
6481 caagccatgc cttgggcttg agcgcttcgg tcttctgtgc cccgcccttc tccggggtct
6541 tgccgttgta ccgcttgaac cactgagcgg cgggccgctc gatgccgtca ttgatccgct
6601 cggagatcat caggtggcag tgcgggttct cgccgccacc ggcatggatg gccagcgtat
6661 acggcaggcg ctcggcaccg gtcaggtgct gggcgaactc ggacgccagc gccttctgct
6721 ggtcgagggt cagctcgacc ggcagggcaa attcgacctc cttgaacagc cgcccattgg
6781 cgcgttcata caggtcggca gcatcccagt agtcggcggg ccgctcgacg aactccggca
6841 tgtgcccgga ttcggcgtgc aagacttcat ccatgtcgcg ggcatacttg ccttcgcgct
6901 ggatgtagtc ggccttggcc ctggccgatt ggccgcccga cctgctgccg gttttcgccg
6961 taaggtgata aatcgccatg ctgcctcgct gttgcttttg cttttcggct ccatgcaatg
7021 gccctcggag agcgcaccgc ccgaagggtg gccgttaggc cagtttctcg aagagaaacc
7081 ggtaagtgcg ccctccccta caaagtaggg tcgggattgc cgccgctgtg cctccatgat
7141 agcctacgag acagcacatt aacaatgggg tgtcaagatg gttaagggga gcaacaaggc
7201 ggcggatcgg ctggccaagc tcgaagaaca acgagcgcga atcaatgccg aaattcagcg
7261 ggtgcgggca agggaacagc agcaagagcg caagaacgaa acaaggcgca aggtgctggt
7321 gggggccatg attttggcca aggtgaacag cagcgagtgg ccggaggatc ggctcatggc
7381 ggcaatggat gcgtaccttg aacgcgacca cgaccgcgcc ttgttcggtc tgccgccacg
7441 ccagaaggat gagccgggct gaatgatcga ccgagacagg ccctgcgggg ctgcacacgc
7501 gcccccaccc ttcgggtagg gggaaaggcc gctaaagcgg ctaaaagcgc tccagcgtat
7561 ttctgcgggg tttggtgtgg ggtttagcgg gctttgcccg cctttccccc tgccgcgcag
7621 cggtggggcg gtgtgtagcc tagcgcagcg aatagaccag ctatccggcc tctggccggg
7681 catattgggc aagggcagca gcgccccaca agggcgctga taaccgcgcc tagtggatta
7741 ttcttagata atcatggatg gatttttcca acaccccgcc agcccccgcc cctgctgggt
7801 ttgcaggttt gggggcgtga cagttattgc aggggttcgt gacagttatt gcaggggggc
7861 gtgacagtta ttgcaggggt tcgtgacagt tagtacggga gtgacgggca ctggctggca
7921 atgtctagca acggcaggca tttcggctga gggtaaaaga actttccgct aagcgataga
7981 ctgtatgtaa acacagtat
//
