质粒名称: | pCDF-NtXR1AtR2B2NtS1 |
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出品公司: | addgene |
目录编号: | 160879 |
存储实验室: | Maureen Hanson |
载体骨架基本信息 |
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载体名称: | pCDF-1b |
完整质粒大小: | 7738 bp |
载体类型: | 细菌细胞表达 |
载体抗性: | Spectinomycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | bcS-S1, rbcX, raf1, raf2, bsd2 |
物种来源: | A. thaliana (mustard weed); Nicotiana tabacum |
启动子: | rbcS-S1: pBAD, chaperones: T7 |
克隆方法: | 限制性内切酶 |
5’克隆位点: | 未知(未破坏) |
文献引用方法 |
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材料和方法部分: | pCDF-NtXR1AtR2B2NtS1 was a gift from Maureen Hanson (Addgene plasmid # 160879 ; http://n2t.net/addgene:160879 ; RRID:Addgene_160879) |
参考文献部分: | Small subunits can determine enzyme kinetics of tobacco Rubisco expressed in Escherichia coli. Lin MT, Stone WD, Chaudhar Vi, Hanson MR. Nat Plants 10.1038/s41477-020-00761-5 |
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如果 LOCUS pCDF-NtXR1AtR2B2NtS1 7738 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of N. tabacum Rubisco small subunit (rbcS-S1), Rubisco chaperone protiens rbcX, raf1 and A. thaliana Rubisco chaperone proteins raf2, and bsd2 in E. coli.. ACCESSION . VERSION . KEYWORDS pCDF-NtXR1AtR2B2NtS1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7738) AUTHORS Lin MT, Stone WD, Chaudhar Vi, Hanson MR TITLE Small subunits can determine enzyme kinetics of tobacco Rubisco expressed in Escherichia coli JOURNAL Nat Plants REFERENCE 2 (bases 1 to 7738) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7738 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 286..1024 /direction=RIGHT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." promoter 1072..1163 /gene="bla" /label=AmpR promoter CDS 1164..1955 /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" promoter 2069..2353 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 2276..2295 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 2380..2402 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" terminator 2884..2931 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(2983..3027) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" terminator 3187..3234 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(3302..3331) /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="ASMTGGQQMG" RBS 3588..3610 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 3625..3649 /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)." promoter complement(3650..3668) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator 3835..3882 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(3950..3979) /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="ASMTGGQQMG" RBS 4506..4528 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 4543..4567 /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)." promoter complement(4568..4586) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5939..5963 /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)." promoter complement(5964..5982) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 6565..6589 /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)." promoter complement(6590..6608) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 6654..6731 /gene="lacI" /label=lacI promoter CDS 6732..76 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(6751..6770) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact 61 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg 121 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg 181 gcatgactaa catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt 241 tgaagagata aattgcactg aaatctagag cggttcagta gaaaagatca aaggatcttc 301 ttgagatcct ttttttctgc gcgtaatctt ttgccctgta aacgaaaaaa ccacctgggg 361 aggtggtttg atcgaaggtt aagtcagttg gggaactgct taaccgtggt aactggcttt 421 cgcagagcac agcaaccaaa tctgtccttc cagtgtagcc ggactttggc gcacacttca 481 agagcaaccg cgtgtttagc taaacaaatc ctctgcgaac tcccagttac caatggctgc 541 tgccagtggc gttttaccgt gcttttccgg gttggactca agtgaacagt taccggataa 601 ggcgcagcag tcgggctgaa cggggagttc ttgcttacag cccagcttgg agcgaacgac 661 ctacaccgag ccgagatacc agtgtgtgag ctatgagaaa gcgccacact tcccgtaagg 721 gagaaaggcg gaacaggtat ccggtaaacg gcagggtcgg aacaggagag cgcaagaggg 781 agcgacccgc cggaaacggt ggggatcttt aagtcctgtc gggtttcgcc cgtactgtca 841 gattcatggt tgagcctcac ggctcccaca gatgcaccgg aaaagcgtct gtttatgtga 901 actctggcag gagggcggag cctatggaaa aacgccaccg gcgcggccct gctgttttgc 961 ctcacatgtt agtcccctgc ttatccacgg aatctgtggg taactttgta tgtgtccgca 1021 gcgcccgccg cagtctcacg cccggagcgt agcgaccgag tgagctagct atttgtttat 1081 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 1141 aataatattg aaaaaggaag agtatgaggg aagcggtgat cgccgaagta tcgactcaac 1201 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt 1261 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta 1321 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa 1381 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc 1441 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc 1501 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta 1561 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac 1621 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat 1681 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca 1741 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa 1801 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg 1861 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg 1921 tgaaaggcga gatcaccaag gtagtcggca aataatgtct aacaattcgt tcaagccgag 1981 gggccgcaag atccggccac gatgacccgg tcgtcggttc agggcagggt cgttaaatag 2041 ccgcttatgt ctattgctgg tttaccggaa gaaaccaatt gtccatattg catcagacat 2101 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt 2161 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc 2221 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc 2281 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct 2341 actgtttctc catacccgtt tttttgggct agaaataatt ttgtttaact ttaagaagga 2401 gatatacata cccatgcagg tgtggccacc aattaacaag aagaagtacg agactctctc 2461 ataccttcct gatttgagcc aggagcaatt gcttagtgaa gttgagtacc ttttgaaaaa 2521 tggatgggtt ccttgcttgg aattcgagac tgagcacgga tttgtctatc gtgaaaacaa 2581 caagtcacca ggatactatg atggcagata ctggaccatg tggaagctac ctatgttcgg 2641 atgcactgat gccacccaag tgttggctga ggtagaagag gcgaagaagg catacccaca 2701 ggcctggatc cgtatcattg gattcgacaa cgtgcgtcaa gtgcagtgca tcagtttcat 2761 tgcctacaag cctgaaggat attaagtcgc aaaaaacccc gcttcggcgg ggttttttcg 2821 ctccggttta ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggttt 2881 cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc 2941 agcggtggca gcagcctagg ttaattaagc tgcgctagta gacgagtcca tgtgctggcg 3001 ttcaaatttc gcagcagcgg tttctttacc agactcgagg gtaccgacgt cagcgatcgc 3061 gtggccggcc gatatccaat tccgcatgcg ctagcacctc cactacaatt gaccgctggt 3121 accgcttggt tatgccggta ctgccgggcc tcttgcggga tatccggata tagttcctcc 3181 tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctagttattg 3241 ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag ccggatctgc 3301 gacccatttg ctgtccacca gtcatgctag cctactcatc gaaggtgctc aagaacccac 3361 caatgaaacc agctccattg cagtccccac acaacacctc ctttttacct ctgcaaagcc 3421 aacagagtgc accagcttta aactgaccat tgaaatgatc aatcaaattc acaccacctc 3481 ctttgcattg agaacatgcc acacaacctt ccccttcaca atttgcacaa actaaactgt 3541 ttggtttagt gccctgagga ttattgtttg cggccatcat atgtatatct ccttcttaaa 3601 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag 3661 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc 3721 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacactag tgcttggtta 3781 tgccggtact gccgggcctc ttgcgggata tccggatata gttcctcctt tcagcaaaaa 3841 acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc 3901 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatccgcga cccatttgct 3961 gtccaccagt catgctagct cacgcccaag ctcttttcct aggggaaaga tcagaagtct 4021 tgatatcatc tattttagcc gccattatga aatcattcat gctcaaccct ccgatagaag 4081 aggtaaatag ttcagctcga acttgggtag gactttccaa gtgaagagaa gggtaatgac 4141 cagaagcttc agcaacctta tggatcctgt ttatgagttc aacaccgcac ccaaaatccc 4201 tcactttcca catacacctt attttcagac cacccgcttc attatccact atactccatc 4261 ccaaaacctt gtggaggaga gttctggcat catcgccgga caaaggaggc tgtgaagacg 4321 aaacagggga acactgaagc tgggagagag acaaaacaga tgcgtttggt atcaaaatct 4381 tgtgctcagt gctttggcat cccaacactt tatctccgaa ttgactcgct atctcctccg 4441 ggtaagggtc acgagctccg aagtcaccaa gaaaatcctg cgccagatta gacatcatat 4501 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca 4561 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc 4621 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc 4681 gacctgcagt ctagaaagct aagcttcttg gtctttaatc ccaatcttct tcacctaatt 4741 gttcttcatc ttcccacctt ggtggcctaa ctactataag cacaattcct aagctctgtt 4801 caacccctct tttcttcaat gttgaccctc tttccacttt caatccctcc tcattgctcc 4861 cgtctccacc atcaaccacc aagtaaaacc cgtcctccga cgccacttct gtccttcccc 4921 tatccgccac taccaaaacc ggctcttgct tgtacttact cttctccctc cacggcaaaa 4981 agttcccgct cttgaacgac actgcaactc cacctctctc taaccccgcg ataggttgcc 5041 atccgggcaa aacaacccat ctcctccaat ccttctccgc ctccactata ccaaaatccc 5101 ctacgccacc acattcccac ggcgccgcct ccacctccac gtcccttccc tccgccttac 5161 aaacaggcaa aacgactaca atcgtagact cagctacctc ccctaacttc attctcacca 5221 aaggcactgt caccccatca tcaagtacac tctccttagc aacctcacgt ccttctagct 5281 ccatcacaag caaattcctc gcactttcag actcaacaac ttctaatgcc ttctccattg 5341 acgacctcct caaatcctca gaagctgccg cagcatattc ttgagctaat ctaaagtaca 5401 taaaagctaa acaatcacca ggagaatcac cattaaactt gtcccaacca taatccacac 5461 gtctacgcgg ataatcttta attgctctag ctaaatcttg agcttgtttt gcatcaaacc 5521 cattcttcac taaaaatgtt gctgcatcag ttcgttgcct agcactaagc aaacgaattt 5581 catataataa ttcagcacct ccagattcaa aatacgacaa cgtttcttca tctaaagcag 5641 cagactcaac aagagtatca cgaacttggg cagcaacaac caaccgattc tgttcaactc 5701 cagtaaggcc agtgatttcc tcaagagttg aaggagtaaa cccttcactt gttaaataag 5761 gtatcaaagg agcatattca taccaaagac caaggcgatt agatagaact tcgagctttg 5821 catttgtgtc aagattgcgg aatttgggtg ggagaggtgg aggaggaggt ctaaaaggct 5881 gataaagttg ttgcatatgt atatctcctt cttatactta actaatatac taagatgggg 5941 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgat tatgcggccg 6001 tgtacaatac gattactttc tgttcgactt aagcattatg cggccgcact gggcgcgcct 6061 cacccaactc tctccgagtt cgtgtcatct gatggccaaa tcacagtttc catgagccgc 6121 tcacgcatca cttccacatt ctgatcagat attcggttgt agatctcctc gtgatcacat 6181 ttttttatcc atttcgcata cagactgagc cgtgttacca tcactttttc agcgagttct 6241 tgcttctcct tagcaaggtt gcgaataaaa cgcttcccat ccctaagctt atttactgta 6301 acataatcat tgaaccattg atattgtgga gagtccattt cattaagctg gttcaaaata 6361 gtcctcactg ctttaaatgt gaagtagtta agaatttgcg ttttgatatc atcaaagcca 6421 tattcatact gtccacctaa atcactgaca accacaagat tcctcctact ctttttctcc 6481 ttctttaaat tcttgtaatt acccttaaat gcttttgcca tggtatatct ccttattaaa 6541 gttaaacaaa attatttcta caggggaatt gttatccgct cacaattccc ctatagtgag 6601 tcgtattaat ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca 6661 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca 6721 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt 6781 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa 6841 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg 6901 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt 6961 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt 7021 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc 7081 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg 7141 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca 7201 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat 7261 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc 7321 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg 7381 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca 7441 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca 7501 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg 7561 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg 7621 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc 7681 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaac //
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