质粒名称: | pET30a-Tn5 |
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出品公司: | 爱迪基因 |
目录编号: | 127917 |
存储实验室: | Silin Zhong |
载体骨架基本信息 |
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载体名称: | pET30a |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
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: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Tn5 |
物种来源: | -- |
插入大小: | -- |
基因突变: | E54K, L372P |
GenBank编号 | -- |
启动子 | T7 |
标签/融合蛋白: | Mxe intein - Chitin-binding domain(插入端的C末端) |
克隆方法: | Gibson克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | T7 |
3' 测序引物 | T7ter |
文献引用方法 |
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材料和方法部分: | pET30a-Tn5 was a gift from Silin Zhong (Addgene plasmid # 127917 ; http://n2t.net/addgene:127917 ; RRID:Addgene_127917) |
买家导航
pET30aTn5载体基本信息
pET30aTn5载体图谱质粒图谱和多克隆位点信息
pET30aTn5载体简介
pET30aTn5载体序列
LOCUS pET30a-Tn5 7445 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Prokaryotic expression of the unmodified Tn5 for Illumina sequencing library preperation.. ACCESSION . VERSION . KEYWORDS pET30a-Tn5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7445) TITLE Zhong Lab plasmids REFERENCE 2 (bases 1 to 7445) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7445 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 23..478 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(110..129) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 320..341 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(570..1385) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 1293..1312 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1377..1396 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 1507..2095 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1996..2015 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2249..2266 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2281..2420 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2346..2365) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2506..2528 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2522..2713) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3522..4604) /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 4566..4585 /label=LacI-R /note="LacI, reverse primer" promoter complement(4605..4682) /gene="lacI" /label=lacI promoter primer_bind 4888..4907 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 4995..5014 /label=T7 /note="T7 promoter, forward primer" promoter 4995..5013 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5014..5038 /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)." RBS 5053..5075 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5083..6510 /codon_start=1 /product="transposase from the bacterial Tn5 transposon (Reznikoff, 1993)" /label=Tn5 transposase /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS KLDGFLAAKDLMAQGIKI" CDS 6511..7104 /codon_start=1 /gene="Mycobacterium xenopi gyrA" /product="modified GyrA intein (Southworth et al., 1999)" /label=Mxe GyrA intein /note="N-terminal cleavage of this intein can be induced by thiols." /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA" CDS 7135..7290 /codon_start=1 /gene="Bacillus circulans chiA" /product="chitin binding domain from chitinase A1 (Watanabe et al., 1994)" /label=CBD /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV PALWQLQ" CDS 7300..7317 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(7370..7388) /label=T7 Term /note="T7 terminator, reverse primer" terminator 7384..7431 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 61 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 121 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 181 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 241 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 301 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 361 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 421 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag 481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 541 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac 601 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat 661 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg 721 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta 781 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc 841 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca 901 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg 961 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca 1021 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg 1081 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc 1141 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg 1201 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat 1261 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa 1321 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg 1381 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa 1441 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 1501 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 1561 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 1621 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca 1681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 1741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 1801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 1861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 1921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 1981 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 2041 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 2101 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 2161 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 2221 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg 2281 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact 2341 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac 2401 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 2461 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 2521 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag 2581 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt 2641 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt 2701 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac 2761 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac 2821 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca 2881 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc 2941 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca 3001 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt 3061 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa 3121 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg 3181 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag 3241 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca 3301 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct 3361 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc 3421 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg 3481 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt 3541 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 3601 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg 3661 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc 3721 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta 3781 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg 3841 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc 3901 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt 3961 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga 4021 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg 4081 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg 4141 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca 4201 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg 4261 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc 4321 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc 4381 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt 4441 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc 4501 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag 4561 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat 4621 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg 4681 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag 4741 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa 4801 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 4861 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 4921 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct 4981 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc 5041 tctagaaata attttgttta actttaagaa ggagatatac atatgattac cagtgcactg 5101 catcgtgcgg cggattgggc gaaaagcgtg ttttctagtg ctgcgctggg tgatccgcgt 5161 cgtaccgcgc gtctggtgaa tgttgcggcg caactggcca aatatagcgg caaaagcatt 5221 accattagca gcgaaggcag caaagccatg caggaaggcg cgtatcgttt tattcgtaat 5281 ccgaacgtga gcgcggaagc gattcgtaaa gcgggtgcca tgcagaccgt gaaactggcc 5341 caggaatttc cggaactgct ggcaattgaa gataccacct ctctgagcta tcgtcatcag 5401 gtggcggaag aactgggcaa actgggtagc attcaggata aaagccgtgg ttggtgggtg 5461 catagcgtgc tgctgctgga agcgaccacc tttcgtaccg tgggcctgct gcatcaagaa 5521 tggtggatgc gtccggatga tccggcggat gcggatgaaa aagaaagcgg caaatggctg 5581 gccgctgctg caacttcgcg tctgagaatg ggcagcatga tgagcaacgt gattgcggtg 5641 tgcgatcgtg aagcggatat tcatgcgtat ctgcaagata aactggccca taacgaacgt 5701 tttgtggtgc gtagcaaaca tccgcgtaaa gatgtggaaa gcggcctgta tctgtatgat 5761 cacctgaaaa accagccgga actgggcggc tatcagatta gcattccgca gaaaggcgtg 5821 gtggataaac gtggcaaacg taaaaaccgt ccggcgcgta aagcgagcct gagcctgcgt 5881 agcggccgta ttaccctgaa acagggcaac attaccctga acgcggtgct ggccgaagaa 5941 attaatccgc cgaaaggcga aaccccgctg aaatggctgc tgctgaccag cgagccggtg 6001 gaaagtctgg cccaagcgct gcgtgtgatt gatatttata cccatcgttg gcgcattgaa 6061 gaatttcaca aagcgtggaa aacgggtgcg ggtgcggaac gtcagcgtat ggaagaaccg 6121 gataacctgg aacgtatggt gagcattctg agctttgtgg cggtgcgtct gctgcaactg 6181 cgtgaatctt ttactccgcc gcaagcactg cgtgcgcagg gcctgctgaa agaagcggaa 6241 cacgttgaaa gccagagcgc ggaaaccgtg ctgaccccgg atgaatgcca actgctgggc 6301 tatctggata aaggcaaacg caaacgcaaa gaaaaagcgg gcagcctgca atgggcgtat 6361 atggcgattg cgcgtctggg cggctttatg gatagcaaac gtaccggcat tgcgagctgg 6421 ggtgcgctgt gggaaggttg ggaagcgctg caaagcaaac tggatggctt tctggccgcg 6481 aaagacctga tggcgcaggg cattaaaatc tgcatcacgg gagatgcact agttgcccta 6541 cccgagggcg agtcggtacg catcgccgac atcgtgccgg gtgcgcggcc caacagtgac 6601 aacgccatcg acctgaaagt ccttgaccgg catggcaatc ccgtgctcgc cgaccggctg 6661 ttccactccg gcgagcatcc ggtgtacacg gtgcgtacgg tcgaaggtct gcgtgtgacg 6721 ggcaccgcga accacccgtt gttgtgtttg gtcgacgtcg ccggggtgcc gaccctgctg 6781 tggaagctga tcgacgaaat caagccgggc gattacgcgg tgattcaacg cagcgcattc 6841 agcgtcgact gtgcaggttt tgcccgcggg aaacccgaat ttgcgcccac aacctacaca 6901 gtcggcgtcc ctggactggt gcgtttcttg gaagcacacc accgagaccc ggacgcccaa 6961 gctatcgccg acgagctgac cgacgggcgg ttctactacg cgaaagtcgc cagtgtcacc 7021 gacgccggcg tgcagccggt gtatagcctt cgtgtcgaca cggcagacca cgcgtttatc 7081 acgaacgggt tcgtcagcca cgctactggc ctcaccggtc tgaactcagg cctcacgaca 7141 aatcctggtg tatccgcttg gcaggtcaac acagcttata ctgcgggaca attggtcaca 7201 tataacggca agacgtataa atgtttgcag ccccacacct ccttggcagg atgggaacca 7261 tccaacgttc ctgccttgtg gcagcttcaa tgactcgagc accaccacca ccaccactga 7321 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 7381 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 7441 ggaac //
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pET30aTn5载体质粒应用举例
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