| 质粒名称: | pcDNA3.4_CEACAM6-His |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 149337 |
| 存储实验室: | James Fleckenstein |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.4_mIgG2-mMxr8a |
| 载体出品公司: | Diamond lab |
| 空载体大小: | 7472 bp |
| 完整质粒大小: | 6465bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | CEACAM6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 858 bp |
| 基因突变: | point mutation (GGA to GCA) to replace 'G-glycine' with 'A-alanine' at the site of lipidation required for GPI anchoring at plasmid position 1772 |
| 启动子: | CMV |
| 标签/融合蛋白: |
polyhistidine (C terminal on insert)
IL-2 signal peptide (N terminal on backbone)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | TAAGTCTTGCACTTGTCACGAATTCGATATCGAAGCTCACTATTGAATCCACG |
| 3’测序引物 | CTCAATGGTGATGGTGATGATGACCGGTTGCAGAGACTGTGATCATCGTG |
|
文献引用方法 |
|
| 材料和方法部分: |
pcDNA3.4_CEACAM6-His was a gift from James Fleckenstein (Addgene plasmid # 149337 ; http://n2t.net/addgene:149337 ; RRID:Addgene_149337)
|
| 参考文献部分: | CEACAMs serve as toxin-stimulated receptors for enterotoxigenic Escherichia coli. Sheikh A, Tumala B, Vickers TJ, Alvarado D, Ciorba MA, Bhuiyan TR, Qadri F, Singer BB, Fleckenstein JM. Proc Natl Acad Sci U S A. 2020 Nov 17;117(46):29055-29062. doi: 10.1073/pnas.2012480117. Epub 2020 Nov 2. 10.1073/pnas.2012480117 PubMed 33139570 |
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 pcDNA3.4_CEACAM6-His 6465 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION for eukaryotic expression and purification of CEACAM6-His6.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.4_CEACAM6-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6465)
AUTHORS Fleckenstein Lab
TITLE CEACAM6 plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6465)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6465
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 850..909
/product="human interleukin-2 signal sequence"
/label=IL-2 signal sequence
CDS 1780..1797
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1820..1837)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1826..2050
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2096..2524
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2183..2202)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2393..2414
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2533..2553)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2538..2867
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2718..2853
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2780..2799
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2934..3728
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2988..3007)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3598..3617
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3904..4025
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3941..3960)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3995..4014
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4074..4090)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4074..4090)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4087..4109)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4098..4114
/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(4122..4152)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4167..4188
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4305..4322)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4476..5064)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4556..4575)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5235..6095)
/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 5858..5877
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6096..6200)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6275..6294)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagttaag cttggtaccg agctcggatc cagtaccctt caccaccatt actgccatct
721 cgatgaaagt ctcgccgtaa tcgaggatta ccactacgct gatgtacaga tccatactgc
781 atgttcattg acgatctaca agctggctag ttaagcttgg taccgagctc ggatccagta
841 cccttcacca tgtacaggat gcaactcctg tcttgcattg cactaagtct tgcacttgtc
901 acgaattcga tatcgaagct cactattgaa tccacgccat tcaatgtcgc agaggggaag
961 gaggttcttc tactcgccca caacctgccc cagaatcgta ttggttacag ctggtacaaa
1021 ggcgaaagag tggatggcaa cagtctaatt gtaggatatg taataggaac tcaacaagct
1081 accccagggc ccgcatacag tggtcgagag acaatatacc ccaatgcatc cctgctgatc
1141 cagaacgtca cccagaatga cacaggattc tataccctac aagtcataaa gtcagatctt
1201 gtgaatgaag aagcaaccgg acagttccat gtatacccgg agctgcccaa gccctccatc
1261 tccagcaaca actccaaccc cgtggaggac aaggatgctg tggccttcac ctgtgaacct
1321 gaggttcaga acacaaccta cctgtggtgg gtaaatggtc agagcctccc ggtcagtccc
1381 aggctgcagc tgtccaatgg caacatgacc ctcactctac tcagcgtcaa aaggaacgat
1441 gcaggatcct atgaatgtga aatacagaac ccagcgagtg ccaaccgcag tgacccagtc
1501 accctgaatg tcctctatgg cccagatgtc cccaccattt ccccctcaaa ggccaattac
1561 cgtccagggg aaaatctgaa cctctcctgc cacgcagcct ctaacccacc tgcacagtac
1621 tcttggttta tcaatgggac gttccagcaa tccacacaag agctctttat ccccaacatc
1681 actgtgaata atagcggatc ctatatgtgc caagcccata actcagccac tggcctcaat
1741 aggaccacag tcacgatgat cacagtctct gcaaccggtc atcatcacca tcaccattga
1801 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
1861 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
1921 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
1981 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
2041 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
2101 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
2161 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
2221 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
2281 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
2341 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
2401 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
2461 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
2521 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
2881 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
2941 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
3001 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
3061 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
3121 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
3181 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
3241 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
3301 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
3361 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
3421 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
3481 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
3541 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
3601 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
3661 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
3721 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
3781 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
3841 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
3901 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
3961 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
4021 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
4081 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
4141 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
4201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
4261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
4321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
4381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
6121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
6181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
6241 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
6301 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
6361 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
6421 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
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