| 质粒名称: | pH6HTC_PTBP1-Halo |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 175302 |
| 存储实验室: | Claudia Kutter |
|
载体骨架基本信息 |
|
| 载体名称: | pH6HTC_minus_cer-region |
| 完整质粒大小: | 5039 bp |
| 载体类型: | 细菌、细胞自由表达 |
| 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: | PTBP1 |
| 物种来源: | H. sapiens (human) |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pH6HTC_PTBP1-Halo was a gift from Claudia Kutter (Addgene plasmid # 175302 ; http://n2t.net/addgene:175302 ; RRID:Addgene_175302) |
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 pH6HTC_PTBP1-Halo 5039 bp ds-DNA circular SYN 07-OCT-2021
DEFINITION Production of HaloTag fusion protein.
ACCESSION .
VERSION .
KEYWORDS pH6HTC_PTBP1-Halo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5039)
TITLE Kutter Lab HaloTag plasmids 2021
REFERENCE 2 (bases 1 to 5039)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 7, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5039
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 55..141
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 233..260
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 342..361
/label=T7
/note="T7 promoter, forward primer"
promoter 342..360
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 400..409
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2026..2046
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="EDLYFQS"
CDS 2062..2946
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
SEIARWLSTLEISG"
CDS 2947..2964
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2975..2993)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2989..3036
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 3265..3369
/gene="bla"
/label=AmpR promoter
CDS 3370..4230
/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(3588..3607)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(4388..4976)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4468..4487)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
361 gtattttaac tttactaagg agcgatcgct tccgaattcg ccaccatgga cggcattgtc
421 ccagatatag ccgttggtac aaagcgggga tctgacgagc ttttctctac ttgtgtcact
481 aacggaccgt ttatcatgag cagcaactcg gcttctgcag caaacggaaa tgacagcaag
541 aagttcaaag gtgacagccg aagtgcaggc gtcccctcta gagtgatcca catccggaag
601 ctccccatcg acgtcacgga gggggaagtc atctccctgg ggctgccctt tgggaaggtc
661 accaacctcc tgatgctgaa ggggaaaaac caggccttca tcgagatgaa cacggaggag
721 gctgccaaca ccatggtgaa ctactacacc tcggtgaccc ctgtgctgcg cggccagccc
781 atctacatcc agttctccaa ccacaaggag ctgaagaccg acagctctcc caaccaggcg
841 cgggcccagg cggccctgca ggcggtgaac tcggtccagt cggggaacct ggccttggct
901 gcctcggcgg cggccgtgga cgcagggatg gcgatggccg ggcagagccc cgtgctcagg
961 atcatcgtgg agaacctctt ctaccctgtg accctggatg tgctgcacca gattttctcc
1021 aagttcggca cagtgttgaa gatcatcacc ttcaccaaga acaaccagtt ccaggccctg
1081 ctgcagtatg cggaccccgt gagcgcccag cacgccaagc tgtcgctgga cgggcagaac
1141 atctacaacg cctgctgcac gctgcgcatc gacttttcca agctcaccag cctcaacgtc
1201 aagtacaaca atgacaagag ccgtgactac acacgcccag acctgccttc cggggacagc
1261 cagccctcgc tggaccagac catggccgcg gccttcggcc tttccgttcc gaacgtccac
1321 ggcgccctgg cccccctggc catcccctcg gcggcggcgg cagctgcggc ggcaggtcgg
1381 atcgccatcc cgggcctggc gggggcagga aattctgtat tgctggtcag caacctcaac
1441 ccagagagag tcacacccca aagcctcttt attcttttcg gcgtctacgg tgacgtgcag
1501 cgcgtgaaga tcctgttcaa taagaaggag aacgccctag tgcagatggc ggacggcaac
1561 caggcccagc tggccatgag ccacctgaac gggcacaagc tgcacgggaa gcccatccgc
1621 atcacgctct cgaagcacca gaacgtgcag ctgccccgcg agggccagga ggaccagggc
1681 ctgaccaagg actacggcaa ctcacccctg caccgcttca agaagccggg ctccaagaac
1741 ttccagaaca tattcccgcc ctcggccacg ctgcacctct ccaacatccc gccctcagtc
1801 tccgaggagg atctcaaggt cctgttttcc agcaatgggg gcgtcgtcaa aggattcaag
1861 ttcttccaga aggaccgcaa gatggcactg atccagatgg gctccgtgga ggaggcggtc
1921 caggccctca ttgacctgca caaccacgac ctcggggaga accaccacct gcgggtctcc
1981 ttctccaagt ccaccatcgg cggctccggc ctcgagccaa ccactgagga tctgtacttt
2041 cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg
2101 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
2161 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
2221 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
2281 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
2341 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
2401 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
2461 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
2521 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
2581 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
2641 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
2701 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
2761 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
2821 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
2881 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt
2941 tccggtcatc atcaccatca ccactaatag tcaaccgctg agcaataact agcataaccc
3001 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccggt
3061 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
3121 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
3181 atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca
3241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
3301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
3361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
3421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
3481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
3541 tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg
3601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
3661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
3721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
3781 acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
3841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
3901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
3961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
4021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
4081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
4141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
4201 ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac
4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggg
//
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