| 质粒名称: | Sfp pet29b C-terminal His Tag |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 75015 |
| 存储实验室: | Michael Burkart |
|
载体骨架基本信息 |
|
| 载体名称: | pet29b |
| 载体出品公司: | Clontech |
| 空载体大小: | 5370bp |
| 完整质粒大小: | 5906bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin |
| 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: | Sfp / 4'-phosphopantetheinyl transferase |
| 物种来源: | B. subtilis |
| GenBank编号 | X63158.1 |
| 启动子: | T7 |
| 标签/融合蛋白: | His Tag (C terminal on backbone) |
| 克隆方法: | 未知 |
| 5’测序引物 | TAATACGACTCACTATAGGG (T7 promoter, forward primer) |
| 3’测序引物 | GCTAGTTATTGCTCAGCGG (T7 terminator, reverse primer) |
|
文献引用方法 |
|
| 材料和方法部分: | Sfp pet29b C-terminal His Tag was a gift from Michael Burkart (Addgene plasmid # 75015 ; http://n2t.net/addgene:75015 ; RRID:Addgene_75015) |
| 参考文献部分: | One-pot chemo-enzymatic synthesis of reporter-modified proteins. Worthington AS, Burkart MD. Org Biomol Chem. 2006 Jan 7;4(1):44-6. Epub 2005 Nov 24. 10.1039/b512735a PubMed 16357994 |
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 Sfp pet29b C-terminal His Tag 5906 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses the PPTase Sfp from B. subtilis; loads CoA analogues onto
ACP.
ACCESSION .
VERSION .
KEYWORDS Sfp pet29b C-terminal His Tag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5906)
AUTHORS Worthington AS, Burkart MD
TITLE One-pot chemo-enzymatic synthesis of reporter-modified proteins.
JOURNAL Org Biomol Chem. 2006 Jan 7;4(1):44-6. Epub 2005 Nov 24.
PUBMED 16357994
REFERENCE 2 (bases 1 to 5906)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5906
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 767..784
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(837..855)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 851..898
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 935..1390
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1022..1041)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1232..1253
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1482..2297)
/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 2205..2224
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2289..2308
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 2419..3007
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2908..2927
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3161..3178
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3193..3332
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3258..3277)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3418..3440
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3434..3625)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4434..5516)
/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 5478..5497
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5517..5594)
/gene="lacI"
/label=lacI promoter
primer_bind 5800..5819
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gaagatttac ggaatttata tggaccgccc
121 gctttcacag gaagaaaatg aacggttcat gactttcata tcacctgaaa aacgggagaa
181 atgccggaga ttttatcata aagaagatgc tcaccgcacc ctgctgggag atgtgctcgt
241 tcgctcagtc ataagcaggc agtatcagtt ggacaaatcc gatatccgct ttagcacgca
301 ggaatacggg aagccgtgca tccctgatct tcccgacgct catttcaaca tttctcactc
361 cggccgctgg gtcattggtg cgtttgattc acagccgatc ggcatagata tcgaaaaaac
421 gaaaccgatc agccttgaga tcgccaagcg cttcttttca aaaacagagt acagcgacct
481 tttagcaaaa gacaaggacg agcagacaga ctatttttat catctatggt caatgaaaga
541 aagctttatc aaacaggaag gcaaaggctt atcgcttccg cttgattcct tttcagtgcg
601 cctgcatcag gacggacaag tatccattga gcttccggac agccattccc catgctatat
661 caaaacgtat gaggtcgatc ccggctacaa aatggctgta tgcgccgcac accctgattt
721 ccccgaggat atcacaatgg tctcgtacga agagctttta ctcgagcacc accaccacca
781 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
841 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
901 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
961 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
1021 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
1081 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
1141 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
1201 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
1261 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
1321 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
1381 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
1441 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
1501 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
1561 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
1621 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
1681 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
1741 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
1801 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
1861 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
1921 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
1981 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
2041 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
2101 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
2161 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
2221 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
2281 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
2341 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
2401 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
2461 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
2521 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
2581 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
2641 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
2701 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
2761 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
2821 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
2881 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
2941 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
3001 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
3061 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
3121 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
3181 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
3241 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
3301 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
3361 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
3421 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
3481 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
3541 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
3601 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
3661 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
3721 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
3781 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
3841 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
3901 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
3961 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
4021 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
4081 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
4141 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
4201 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
4261 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
4321 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
4381 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
4441 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
4501 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
4561 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
4621 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
4681 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
4741 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
4801 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
4861 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
4921 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
4981 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
5041 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
5101 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
5161 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
5221 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
5281 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
5341 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
5401 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
5461 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
5521 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
5581 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
5641 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
5701 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
5761 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
5821 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
5881 cgagatcgat ctcgatcccg cgaaat
//
