| 质粒名称: | pHen2 Avi-tag-linker-p3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119819 |
| 存储实验室: | Alexander Gabibov |
|
载体骨架基本信息 |
|
| 载体名称: | pHen2 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 4563bp |
| 载体类型: | 细菌表达 |
| 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: | linker with Avi-tag and p3 |
| 标签/融合蛋白: | Avi-tag (C terminal on insert) |
|
文献引用方法 |
|
| 材料和方法部分: |
pHen2 Avi-tag-linker-p3 was a gift from Alexander Gabibov (Addgene plasmid # 119819 ; http://n2t.net/addgene:119819 ; RRID:Addgene_119819)
|
| 参考文献部分: | Probing surface membrane receptors using engineered bacteriophage bioconjugates. Lomakin YA, Kaminskaya AN, Stepanov AV, Shmidt AA, Gabibov A, Belogurov AA. Bioconjug Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218. 10.1021/acs.bioconjchem.9b00218 PubMed 31021608 |
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pHen2 Avi-tag-linker-p3载体质粒基本信息
pHen2 Avi-tag-linker-p3质粒图谱载体图谱和pHen2 Avi-tag-linker-p3载体序列质粒序列多克隆位点信息
pHen2 Avi-tag-linker-p3质粒载体简介
pHen2 Avi-tag-linker-p3质粒序列载体序列
LOCUS pHen2 Avi-tag-linker-p3 4563 bp DNA circular SYN 14-APR-2025
DEFINITION For cloning and subsequent expression of peptide fragments fused
with Avi-tag (for biotinylation) and with pIII protein of M13K07
bacteriophage.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4563)
AUTHORS Lomakin YA, Kaminskaya AN, Stepanov AV, Shmidt AA, Gabibov A,
Belogurov AA
TITLE Probing surface membrane receptors using engineered bacteriophage
bioconjugates.
JOURNAL Bioconjug Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218.
PUBMED 31021608
REFERENCE 2 (bases 1 to 4563)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Bioconjug
Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218."
COMMENT Sequence Label: pHen2 Avi-tag-linker-p3
FEATURES Location/Qualifiers
source 1..4563
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 59..103
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
primer_bind complement(1361..1378)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1361..1377)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1370..1392)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1590..2045
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1677..1696)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1887..1908
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2062..2081)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2181..2203
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2241..2259)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2327..2431
/gene="bla"
/label=AmpR promoter
CDS join(2432..2500,2501..3292)
/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 complement(2650..2669)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3463..4051
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3952..3971
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4205..4222
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4339..4360
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(4375..4380,4381..4398,4399..4405)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4413..4429
/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 4418..4440
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4437..4453
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4437..4453
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
sig_peptide join(4505..4563,1..7)
/codon_start=1
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
/translation="MKYLLPTAAAGLLLLAAQPAMA"
ORIGIN
1 catggcctta gctagcagtg gtggaggcgg ttcaggcgga ggtggctctg gcggtagtgg
61 tctgaacgac atcttcgagg ctcagaaaat cgaatggcac gaaagtggtg gaggcggttc
121 aactgttgaa agttgtttag caaaacctca tacagaaaat tcatttacta acgtctggaa
181 agacgacaaa actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg
241 cgttgtggtt tgtactggtg acgaaactca gtgttacggt acatgggttc ctattgggct
301 tgctatccct gaaaatgagg gtggtggctc tgagggtggc ggttctgagg gtggcggttc
361 tgagggtggc ggtactaaac ctcctgagta cggtgataca cctattccgg gctatactta
421 tatcaaccct ctcgacggca cttatccgcc tggtactgag caaaaccccg ctaatcctaa
481 tccttctctt gaggagtctc agcctcttaa tactttcatg tttcagaata ataggttccg
541 aaataggcag ggtgcattaa ctgtttatac gggcactgtt actcaaggca ctgaccccgt
601 taaaacttat taccagtaca ctcctgtatc atcaaaagcc atgtatgacg cttactggaa
661 cggtaaattc agagactgcg ctttccattc tggctttaat gaggatccat tcgtttgtga
721 atatcaaggc caatcgtctg acctgcctca acctcctgtc aatgctggcg gcggctctgg
781 tggtggttct ggtggcggct ctgagggtgg cggctctgag ggtggcggct ctgagggtgg
841 cggttctgag ggtggcggct ctgagggtgg cggttccggt ggcggctccg gttccggtga
901 ttttgattat gaaaaaatgg caaacgctaa taagggggct atgaccgaaa atgccgatga
961 aaacgcgcta cagtctgacg ctaaaggcaa acttgattct gtcgctactg attacggtgc
1021 tgctatcgat ggtttcattg gtgacgtttc cggccttgct aatggtaatg gtgctactgg
1081 tgattttgct ggctctaatt cccaaatggc tcaagtcggt gacggtgata attcaccttt
1141 aatgaataat ttccgtcaat atttaccttc tttgcctcag tcggttgaat gtcgccctta
1201 tgtctttggc gctggtaaac catatgaatt ttctattgat tgtgacaaaa taaacttatt
1261 ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt atgtatgtat tttcgacgtt
1321 tgctaacata ctgcgtaata aggagtctta ataagaattc actggccgtc gttttacaac
1381 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
1441 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
1501 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
1561 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
1621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
1681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
1741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
1801 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
1861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
1921 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
1981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
2041 aattttatgg tgcagtctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
2101 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
2161 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
2221 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
2281 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
2341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
2401 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
2461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
2521 agtaaaagat gctgaagatc agttgggtgc ccgagtgggt tacatcgaac tggatctcaa
2581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
2641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
2701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
2761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
2821 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
2881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
2941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
3001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
3061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
3121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
3181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
3241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
3301 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
3361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
3421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
3481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
3541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
3601 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
3781 aacggggggt tcgtgcatac agcccagctt ggagcgaacg acctacaccg aactgagata
3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
4021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
4081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
4141 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
4201 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
4261 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
4321 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
4381 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
4441 aaacagctat gaccatgatt acgccaagct tgcatgcaaa ttctatttca aggagacagt
4501 cataatgaaa tacctattgc ctacggcagc cgctggattg ttattactcg cggcccagcc
4561 ggc
//
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