质粒名称: | pG1AK-sfGFP |
---|---|
出品公司: | addgene |
目录编号: | 71739 |
存储实验室: | Tom Ellis |
载体骨架基本信息 |
|
载体名称: | pG1AK |
空载体大小: | 4719 bp |
完整质粒大小: | 5650 bp |
载体类型: | 细菌表达,合成生物学;穿梭载体 |
载体抗性: | 氨苄霉素 |
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: | sfGFP/super folder Green Fluorescent Protein |
插入大小: | 720 bp |
启动子: | pRplsWT |
克隆方法: | Gibson Cloning |
3’测序引物 | AGCGGATAACAATTTCACACAGGA |
文献引用方法 |
|
材料和方法部分: | pG1AK-sfGFP was a gift from Tom Ellis (Addgene plasmid # 71739 ; http://n2t.net/addgene:71739 ; RRID:Addgene_71739) |
参考文献部分: | The Geobacillus Plasmid Set: A Modular Toolkit for Thermophile Engineering. Reeve B, Martinez-Klimova E, de Jonghe J, Leak DJ, Ellis T. ACS Synth Biol. 2016 Jul 1. 10.1021/acssynbio.5b00298 PubMed 27332993 |
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 pG1AK-sfGFP 5650 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Modular shuttle vector for Geobacillus and E. coli. ACCESSION . VERSION . KEYWORDS pG1AK-sfGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5650) AUTHORS Reeve B, Martinez-Klimova E, de Jonghe J, Leak DJ, Ellis T TITLE The Geobacillus Plasmid Set: A Modular Toolkit for Thermophile Engineering. JOURNAL ACS Synth Biol. 2016 Jul 1. PUBMED 27332993 REFERENCE 2 (bases 1 to 5650) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..5650 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(14..30) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(14..30) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(27..49) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 38..54 /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(62..92) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(245..262) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(416..1004) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(496..515) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1198..2058) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1821..1840 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2059..2163) /gene="bla" /label=AmpR promoter primer_bind 2231..2249 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" CDS 4874..5587 /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" ORIGIN 1 aaatgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 61 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 1021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 1081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 1141 tctaaagtat atatgagtaa acttggtctg acagttacca atggccggcc ctgacagtta 1201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 1261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 1321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 1381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 1441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 1501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 1561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 1621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 1681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 1741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 1801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 1861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 1921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 1981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 2041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 2101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 2161 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 2221 aacctataaa aataggcgta tcacgaggcc gcggccgcct gtcgagaatt tttggaaaat 2281 agaccttgtc gcgactcgaa gattagtagt acaatagaga caacataaca gaacgggaca 2341 aaatagaaga gcttccgaag cgtgtgtaag tggtgctccc aacaccgttc gccgtactcg 2401 ccctacggca aacacctgct cgaaagctct tgtgttaaca tatttatgcc ttgtcttcat 2461 tttaatgcat agatatgttg aatgcaagag cgaaacgctt ggaaattttc gtttttcctc 2521 cgatgagagc gtgtttgcct gcgtattaat gtacggcaac gcgctctttt gtttgtcccg 2581 aaaaattcgg gagaaacggg aggagaacgg aaatgtacat aatcactagc gaacaattca 2641 cgcaactgaa agaaatcgag agagtcacag atttcgttta taaacgccgt aagattaaaa 2701 atgagctgat ggagcgtatc gccgacgcat taggccgcaa gttccgcaaa gcgcaaaaaa 2761 caacgcgcgt catcgacgaa attgtttggc tcgcaacaga ccgcggtttt tcgttcccag 2821 ggcgcgaaac gcttgccaaa aagttaggcg tgtccttgcc gactgttgat cgtgctactc 2881 gcatgctcaa agactcgggc gaagtcgtcg tttgctatcg tgagaatccg aatagtaacg 2941 gacctaaaac gcctgtattc attttccgga gccatgcaaa tttcgagcgt attgcagccg 3001 ttttaaactt gcgtgacaac gaagctgata aagtagaaaa cgccgaaaag cctacggaat 3061 caagcgattt agcgcgtaaa acggacgcta ccattagttc accagttgaa aaacatgacg 3121 atattaataa attaatagac aatacacatc cgttggataa aattgtaaaa tacgtcacta 3181 ttaaagtgaa cgaagtgcag aagcgtcatc cttacggcat tcgatatcta tccgcataca 3241 taaacaaaac actcgacgac ctaatgcaac gtgcaaaagc gaaggtcaaa gccgaaaaag 3301 ctcgcgccca acaacgcaaa caagccgact atcaaccgac cgcgaccggc ttaatttggt 3361 acgacttcac ggcggacaca agcgccagcc agccgacaag ccagcgcaag ccagcgtttg 3421 acagcgaaga catgtttgcg ccgttattgc aggctgttgc acgaattaag ggcgttgagg 3481 tgtgacacac tcgttttaag gcgtctaaca ggcgcgtaga gacgtcttta agtcggggga 3541 atattaaaaa ttccacgcct cctaaaacgc gatatacgcc gattatgagt ttaaacgaag 3601 attagatgct ataattgtta ttaaaaggat tgaaggatgc ttaggaagac gagttattaa 3661 tagctgaata agaacggtgc tctccaaata ttcttattta gaaaagcaaa tctaaaatta 3721 tctgaaaagg gaatgagaat agtgaatgga ccaataataa tgactagaga agaaagaatg 3781 aagattgttc atgaaattaa ggaacgaata ttggataaat atggggatga tgttaaggct 3841 attggtgttt atggctctct tggtcgtcag actgatgggc cctattcgga tattgagatg 3901 atgtgtgtca tgtcaacaga ggaagcagag ttcagccatg aatggacaac cggtgagtgg 3961 aaggtggaag tgaattttta tagcgaagag attctactag attatgcatc tcaggtggaa 4021 tcagattggc cgcttacaca tggtcaattt ttctctattt tgccgattta tgattcaggt 4081 ggatacttag agaaagtgta tcaaactgct aaatcggtag aagcccaaaa gttccacgat 4141 gcgatttgtg cccttatcgt agaagagctg tttgaatatg caggcaaatg gcgtaatatt 4201 cgtgtgcaag gaccgacaac atttctacca tccttgactg tacaggtagc aatggcaggt 4261 gccatgttga ttggtctgca tcatcgcatc tgttatacga cgagcgcttc ggtcttaact 4321 gaagcagtta agcaatcaga tcttccttca ggttatgacc atctgtgcca gttcgtaatg 4381 tctggtcaac tttccgactc tgagaaactt ctggaatcgc tagagaattt ctggaatggg 4441 attcaggagt ggacagaacg acacggatat atagtggatg tgtcaaaacg cataccattt 4501 tgataaggcg cgcctctacg gaccgagtat tcaacaaacg ggccagtttg ttgaaatcgt 4561 taattaagcg tactagtcct aggctagctg ataccaggtc agtcccggga caggtcgacc 4621 tcgagtacgc ctgcaggaac aatcgttaaa gcggacgttt ttgcgccgcc cggatttgct 4681 tgaaaactac ccgctgacag aaaagcaaaa acgatggatc gaagagtgga aaaaagaaaa 4741 acagtagcta ttgcgcatga tacaagttta tgctactata ttccttgtgc aacttaagcg 4801 atttgcttaa gcgaggaaaa cgatgttccg ctgcaatgat gaaaaagcat tgtctagata 4861 aggagtgatt cgaatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt 4921 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga 4981 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc 5041 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga 5101 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg 5161 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg 5221 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat 5281 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa 5341 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt 5401 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc 5461 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga 5521 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact 5581 gtacaaatga taagagctca atttcgcgcc ccgaaagggg cgtttttttg cgaagcctaa 5641 ggtatcattt //
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇:pPB-AMBRA1-3xFLAG
- 下一篇:Cx43-sfGFP