| 质粒名称: | secreted BirA-Flag |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64395 |
| 存储实验室: | Gavin Wright |
|
载体骨架基本信息 |
|
| 载体名称: | pTT3 |
| 载体出品公司: | Yves Durocher |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6000 bp |
| 完整质粒大小: | 7043 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | BirA,Biotin ligase |
| 物种来源: | E. coli |
| 插入大小: | 1000 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | Flag tag（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Sal1（在克隆过程中被破坏） |
| 3' 克隆位点: | Xba1 (未破坏) |
| 5' 测序引物 | CMV forward |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | secreted BirA-Flag was a gift from Gavin Wright (Addgene plasmid # 64395 ; http://n2t.net/addgene:64395 ; RRID:Addgene_64395) |
| 参考文献部分: | Large-scale screening for novel low-affinity extracellular protein interactions. Bushell KM, Sollner C, Schuster-Boeckler B, Bateman A, Wright GJ. Genome Res. 2008 Apr;18(4):622-30. Epub 2008 Feb 22. 10.1101/gr.7187808 PubMed 18296487 |
买家导航
secreted BirA-Flag载体质粒基本信息
secreted BirA-Flag质粒图谱载体图谱和secreted BirA-Flag载体序列质粒序列多克隆位点信息
secreted BirA-Flag质粒载体简介
secreted BirA-Flag质粒序列载体序列
LOCUS secreted BirA-Flag 7001 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Flag-tagged biotinylation enzyme for co-expression with
bio-proteins.
ACCESSION .
VERSION .
KEYWORDS secreted BirA-Flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7001)
AUTHORS Bushell KM, Sollner C, Schuster-Boeckler B, Bateman A, Wright GJ
TITLE Large-scale screening for novel low-affinity extracellular protein
interactions.
JOURNAL Genome Res. 2008 Apr;18(4):622-30. Epub 2008 Feb 22.
PUBMED 18296487
REFERENCE 2 (bases 1 to 7001)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7001
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 154..533
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 534..737
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 688..708
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 734..758
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 1239..1258
/label=pMT2-F
/note="Synthetic intron, forward primer"
CDS 1362..2321
/codon_start=1
/product="E. coli biotin protein ligase"
/label=BirA
/translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
CDS 2328..2351
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2381..2436
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2435..2454)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4672..4690)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4758..4862
/gene="bla"
/label=AmpR promoter
CDS 4863..5723
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5081..5100)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5894..6482
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6383..6402
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6636..6653
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6770..6791
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6806..6836
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6844..6860
/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)."
primer_bind 6849..6871
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6868..6884
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6868..6884
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
1261 ccactcccag gtccaagttt aaacggatct ctagcgaatt catgggcaga cacctcctgc
1321 tgctcctgtt cagcatcctg tacatgctgc tggcccaggc caaggacaac accgtgcccc
1381 tgaagctgat cgccctgctg gccaacggcg agttccacag cggcgagcag ctgggcgaga
1441 ccctgggcat gagcagagcc gctatcaaca agcacatcca gaccctgaga gactggggcg
1501 tggacgtgtt caccgtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga
1561 acgccaagca gatcctgggc cagctggacg gcggaagcgt ggccgtgctg cccgtgatcg
1621 acagcaccaa ccagtacctg ctggacagaa tcggcgagct gaagagcggc gacgcctgca
1681 tcgccgagta ccagcaggcc ggcagaggca gaagaggcag aaagtggttc agcccctttg
1741 gcgccaacct gtacctgagc atgttctgga gactggagca gggccccgca gccgcaatcg
1801 gcctgagcct ggtgatcggc atcgtgatgg ccgaggtgct gagaaagctg ggcgccgaca
1861 aggtgagagt gaagtggccc aacgacctgt acctgcagga cagaaagctg gccggcatcc
1921 tggtggagct gaccggcaag accggcgacg ccgcacagat cgtgatcggc gccggcatca
1981 acatggccat gagaagagtg gaggaaagcg tggtgaacca gggctggatc accctgcagg
2041 aggccggcat caacctggac agaaacaccc tggcagccat gctgatcaga gagctgagag
2101 ccgcactgga gctgttcgag caggagggcc tggctcccta cctgagcaga tgggagaagc
2161 tggacaactt catcaacaga cccgtgaagc tgatcatcgg cgacaaggag atcttcggca
2221 tcagcagagg catcgacaag cagggcgccc tgctgctgga gcaggacggc atcatcaagc
2281 cctggatggg aggcgagatc agcctgagaa gcgccgagaa gggccccgac tacaaggacg
2341 atgatgacaa gtgaggatcc cccgacctcg acctctggct aataaaggaa atttattttc
2401 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
2461 aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac
2521 gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct
2581 tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac
2641 caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg
2701 ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac
2761 attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc
2821 tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt
2881 agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg
2941 ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc
3001 ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa
3061 aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat
3121 ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat
3181 caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta
3241 gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa
3301 ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc
3361 tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa
3421 acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg
3481 atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata
3541 ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac
3601 tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc
3661 tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa
3721 gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc
3781 cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa
3841 ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca
3901 taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca
3961 cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg
4021 agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca
4081 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
4141 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
4201 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
4261 tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta
4321 tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg
4381 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg
4441 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
4501 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
4561 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
4621 ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga
4681 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
4741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
4801 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
4861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
4921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
4981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
5041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
5101 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
5161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
5221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
5281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
5341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
5401 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
5461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
5521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
5581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
5641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
5701 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
5761 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
5821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
5881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
5941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
6001 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
6061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
6121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
6181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
6241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
6301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
6361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
6421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
6481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
6541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
6601 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
6661 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
6721 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
6781 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
6841 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
6901 tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
6961 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg a
//
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