| 质粒名称: | pHL-sec |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99845 |
| 存储实验室: | Edith Yvonne Jones |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG.I-SceI |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | ephrin b2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | His（插接件上的C端子） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pHL-sec was a gift from Edith Yvonne Jones (Addgene plasmid # 99845 ; http://n2t.net/addgene:99845 ; RRID:Addgene_99845) |
| 参考文献部分: | A time- and cost-efficient system for high-level protein production in mammalian cells. Aricescu AR, Lu W, Jones EY. Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50. Epub 2006 Sep 19. 10.1107/S0907444906029799 PubMed 17001101 |
买家导航
pHLsec载体基本信息
pHLsec载体图谱质粒图谱和多克隆位点信息
pHLsec载体简介
pHLsec载体序列
LOCUS pHL-sec 5067 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression with secretion signal sequence.
ACCESSION .
VERSION .
KEYWORDS pHL-sec
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5067)
AUTHORS Aricescu AR, Lu W, Jones EY
TITLE A time- and cost-efficient system for high-level protein production
in mammalian cells.
JOURNAL Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50.
Epub 2006 Sep 19.
PUBMED 17001101
REFERENCE 2 (bases 1 to 5067)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5067
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 385..662
/label=chicken beta-actin promoter
intron 663..1679
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1603..1625
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1687..1706
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1761..1770
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2292..2309
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2421..2440)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2486..2541
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2540..2559)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2902..2918)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2902..2918)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2915..2937)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2926..2942
/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)."
promoter complement(2950..2980)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2995..3016
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3145..3162)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3316..3904)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3396..3415)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4075..4935)
/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 4698..4717
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4936..5040)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc
1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggtt
1861 cgatagtttt agagcctatc tattggaatt cctcgaactc caaatttcta cctggacaag
1921 gactggtact atacccacag ataggagaca aattggatat tatttgcccc aaagtggact
1981 ctaaaactgt tggccagtat gaatattata aagtttatat ggttgataaa gaccaagcag
2041 acagatgcac tattaagaag gaaaataccc ctctcctcaa ctgtgccaaa ccggaccaag
2101 atatcaaatt caccatcaag tttcaagaat tcagccctaa cctctggggt ctagaatttc
2161 agaagaacaa agattattac attatatcta catcaaatgg gtctttggag ggcctggata
2221 accaggaggg aggggtgtgc cagacaagag ccatgaagat cctcatgaaa gttggacaag
2281 atggtaccaa gcaccaccat caccatcact aatgatcact cgagactagt atcgcgataa
2341 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
2401 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
2461 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
2521 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
2581 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
2641 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
2701 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
2761 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
2821 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
2881 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
2941 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
3001 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
3061 gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
3121 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
3181 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
3241 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
3301 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
3361 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
3421 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
3481 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
3541 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
3601 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
3661 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
3721 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
3781 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
3841 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
3901 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
3961 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4021 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
4081 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
4141 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
4201 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
4261 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
4321 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
4381 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
4441 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
4501 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
4561 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
4621 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
4681 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
4741 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
4801 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
4861 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
4921 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
4981 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
5041 cacatttccc cgaaaagtgc cacctgg
//
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