| 质粒名称: | pJet1.2 sfGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 117123 |
| 存储实验室: | Stephan Gruber |
|
载体骨架基本信息 |
|
| 载体名称: | pJet1.2 |
| 载体出品公司: | Thermo scientific |
| 空载体大小: | 2974 bp |
| 完整质粒大小: | 3745 bp |
| 载体类型: |
Golden gate donor vector for gene targeting in Bacillus subtilis
|
| 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: | Superfolder GFP/sfGFP |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 750bp |
| GenBank编号 | CAA65278 |
| 标签/融合蛋白: | sfGFP (C terminal on insert) |
| 克隆方法: | 未知 |
| 5’测序引物 | CGACTCACTATAGGGAGAGCGGC |
| 3’测序引物 | AAGAACATCGATTTTCCATGGCAG |
|
文献引用方法 |
|
| 材料和方法部分: |
pJet1.2 sfGFP was a gift from Stephan Gruber (Addgene plasmid # 117123 ; http://n2t.net/addgene:117123 ; RRID:Addgene_117123)
|
| 参考文献部分: | High-Throughput Allelic Replacement Screening in Bacillus subtilis. Diebold-Durand ML, Burmann F, Gruber S. Methods Mol Biol. 2019;2004:49-61. doi: 10.1007/978-1-4939-9520-2_5. 10.1007/978-1-4939-9520-2_5 PubMed 31147909 |
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 pJet1.2 sfGFP 3745 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION C-terminal GFP tag.
ACCESSION .
VERSION .
KEYWORDS pJet1.2 sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3745)
AUTHORS Diebold-Durand ML, Burmann F, Gruber S
TITLE High-Throughput Allelic Replacement Screening in Bacillus subtilis.
JOURNAL Methods Mol Biol. 2019;2004:49-61. doi: 10.1007/978-1-4939-9520-2_5.
PUBMED 31147909
REFERENCE 2 (bases 1 to 3745)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3745
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 305..324
/label=T7
/note="T7 promoter, forward primer"
promoter 305..323
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(383..1090)
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="KGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFI
CTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANF
KIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
primer_bind complement(423..443)
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind 1038..1066
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
promoter complement(1577..1607)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 1622..1643
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1763..1780)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1934..2522)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2014..2033)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2693..3553)
/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 3316..3335
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3554..3658)
/gene="bla"
/label=AmpR promoter
primer_bind 3726..3744
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
361 ttcagcaaga tggtctcgct tatttgtaga gctcatccat gccatgtgta atcccagcag
421 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgaaagga
481 cagattgtgt cgacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag
541 tattttgttg ataatggtct gctagttgaa cggaaccatc ttcaacgttg tggcgaattt
601 tgaagttagc tttgattcca ttcttttgtt tgtctgccgt gatgtataca ttgtgtgagt
661 taaagttgta ctcgagtttg tgtccaagaa tgtttccatc ttctttaaaa tcaataccct
721 ttaactcgat acgattaaca agggtatcac cttcaaactt gacttcagca cgcgtcttgt
781 aggtcccgtc atctttgaaa gatatagtgc gttcctgtac ataaccttcg ggcatggcac
841 tcttgaaaaa gtcatgccgt ttcatgtgat ccggataacg ggaaaagcat tgaacaccat
901 aggtcagagt agtgacaagt gttggccacg gaacaggtag ttttccagta gtgcaaataa
961 atttaagggt gagttttccg tttgtagcat caccttcacc ctctccacgg acagaaaatt
1021 tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca gtgaaaagtt
1081 cttctccttt gcctccagat ccgcctgagc ctccgctgcc tgatccgcct ccgctcgaga
1141 ccatctttct agaagatctc ctacaatatt ctcagctgcc atggaaaatc gatgttcttc
1201 ttttattctc tcaagatttt caggctgtat attaaaactt atattaagaa ctatgctaac
1261 cacctcatca ggaaccgttg taggtggcgt gggttttctt ggcaatcgac tctcatgaaa
1321 actacgagct aaatattcaa tatgttcctc ttgaccaact ttattctgca ttttttttga
1381 acgaggttta gagcaagctt caggaaactg agacaggaat tttattaaaa atttaaattt
1441 tgaagaaagt tcagggttaa tagcatccat tttttgcttt gcaagttcct cagcattctt
1501 aacaaaagac gtctcttttg acatgtttaa agtttaaacc tcctgtgtga aattattatc
1561 cgctcataat tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
1621 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gccaattgct ttccagtcgg
1681 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
1741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
1801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
1861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
1921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
1981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
2041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
2101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
2161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
2221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
2281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
2341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
2401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
2461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
2521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
2581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
2641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
2701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
2761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
2821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
2881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
2941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
3001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
3061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
3121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
3181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
3241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
3301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
3361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
3421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
3481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
3541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
3601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
3661 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
3721 ataaaaatag gcgtatcacg aggcc
//
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