| 质粒名称: | pc DNA FRT TO- APPSwed/Ind |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 114194 |
| 存储实验室: | Aleksandra Radenovic |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA FRT TO |
| 载体出品公司: | Thermo Fisher Scientific |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳细胞表达 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Amyloid-Precursor-Protein,APP |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2088 bp |
| 基因突变: | Popular mouse model, overexpress 2 mutations: Swedish + Indiana :Indiana= V717F (GTG---TTC); Swedish= KM670/671NL (AAG ATG---AAT CTG) |
| 启动子 | CMV |
| 克隆方法: | Gibson克隆 |
|
文献引用方法 |
|
| 材料和方法部分: | pc DNA FRT TO- APPSwed/Ind was a gift from Aleksandra Radenovic (Addgene plasmid # 114194 ; http://n2t.net/addgene:114194 ; RRID:Addgene_114194) |
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 pc DNA FRT TO- APPSwed/Ind 7225 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Creation of a stable cell line expressing APP mutation (AD in vitro
study).
ACCESSION .
VERSION .
KEYWORDS pc DNA FRT TO- APPSwed/Ind
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7225)
TITLE Radenovic Lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7225)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7225
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 97..476
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 477..680
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 631..651
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 682..700
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 703..721
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 731..755
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS complement(1217..1228)
/codon_start=1
/gene="cspA"
/product="translation enhancing element for E. coli (Qing
et al., 2004)"
/label=TEE
/translation="NHKV"
CDS 1708..1728
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
primer_bind complement(3039..3056)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3045..3269
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3402..3421)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 3553..3600
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 3608..4630
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 4760..4881
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4797..4816)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4851..4870
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4930..4946)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4930..4946)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4943..4965)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4954..4970
/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(4978..5008)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5023..5044
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5161..5178)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5332..5920)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5412..5431)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6091..6951)
/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 6714..6733
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6952..7056)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7131..7150)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
61 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
661 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag
721 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
781 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta
841 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca tgctgcccgg
901 tttggcactg ctcctgctgg ccgcctggac ggctcgggcg ctggaggtac ccactgatgg
961 taatgctggc ctgctggctg aaccccagat tgccatgttc tgtggcagac tgaacatgca
1021 catgaatgtc cagaatggga agtgggattc agatccatca gggaccaaaa cctgcattga
1081 taccaaggaa ggcatcctgc agtattgcca agaagtctac cctgaactgc agatcaccaa
1141 tgtggtagaa gccaaccaac cagtgaccat ccagaactgg tgcaagcggg gccgcaagca
1201 gtgcaagacc catccccact ttgtgattcc ctaccgctgc ttagttggtg agtttgtaag
1261 tgatgccctt ctcgttcctg acaagtgcaa attcttacac caggagagga tggatgtttg
1321 cgaaactcat cttcactggc acaccgtcgc caaagagaca tgcagtgaga agagtaccaa
1381 cttgcatgac tacggcatgt tgctgccctg cggaattgac aagttccgag gggtagagtt
1441 tgtgtgttgc ccactggctg aagaaagtga caatgtggat tctgctgatg cggaggagga
1501 tgactcggat gtctggtggg gcggagcaga cacagactat gcagatggga gtgaagacaa
1561 agtagtagaa gtagcagagg aggaagaagt ggctgaggtg gaagaagaag aagccgatga
1621 tgacgaggac gatgaggatg gtgatgaggt agaggaagag gctgaggaac cctacgaaga
1681 agccacagag agaaccacca gcattgccac caccaccacc accaccacag agtctgtgga
1741 agaggtggtt cgagttccta caacagcagc cagtacccct gatgccgttg acaagtatct
1801 cgagacacct ggggatgaga atgaacatgc ccatttccag aaagccaaag agaggcttga
1861 ggccaagcac cgagagagaa tgtcccaggt catgagagaa tgggaagagg cagaacgtca
1921 agcaaagaac ttgcctaaag ctgataagaa ggcagttatc cagcatttcc aggagaaagt
1981 ggaatctttg gaacaggaag cagccaacga gagacagcag ctggtggaga cacacatggc
2041 cagagtggaa gccatgctca atgaccgccg ccgcctggcc ctggagaact acatcaccgc
2101 tctgcaggct gttcctcctc ggcctcgtca cgtgttcaat atgctaaaga agtatgtccg
2161 cgcagaacag aaggacagac agcacaccct aaagcatttc gagcatgtgc gcatggtgga
2221 tcccaagaaa gccgctcaga tccggtccca ggttatgaca cacctccgtg tgatttatga
2281 gcgcatgaat cagtctctct ccctgctcta caacgtgcct gcagtggccg aggagattca
2341 ggatgaagtt gatgagctgc ttcagaaaga gcaaaactat tcagatgacg tcttggccaa
2401 catgattagt gaaccaagga tcagttacgg aaacgatgct ctcatgccat ctttgaccga
2461 aacgaaaacc accgtggagc tccttcccgt gaatggagag ttcagcctgg acgatctcca
2521 gccgtggcat tcttttgggg ctgactctgt gccagccaac acagaaaacg aagttgagcc
2581 tgttgatgcc cgccctgctg ccgaccgagg actgaccact cgaccaggtt ctgggttgac
2641 aaatatcaag acggaggaga tctctgaagt gaatctggat gcagaattcc gacatgactc
2701 aggatatgaa gttcatcatc aaaaattggt gttctttgca gaagatgtgg gttcaaacaa
2761 aggtgcaatc attggactca tggtgggcgg tgttgtcata gcgacagtga tcttcatcac
2821 cttggtgatg ctgaagaaga aacagtacac atccattcat catggtgtgg tggaggttga
2881 cgccgctgtc accccagagg agcgccacct gtccaagatg cagcagaacg gctacgaaaa
2941 tccaacctac aagttctttg agcagatgca gaactagaga tatccagcac agtggcggcc
3001 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
3061 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
3121 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
3181 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
3241 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
3301 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
3361 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
3421 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
3481 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
3541 ttcacgtacc tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc
3601 cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
3661 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
3721 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
3781 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
3841 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
3901 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
3961 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
4021 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
4081 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
4141 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
4201 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
4261 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
4321 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
4381 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
4441 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
4501 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
4561 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
4621 aaaggaatag cacgtactac gagatttcga ttccaccgcc gccttctatg aaaggttggg
4681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
4741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
4801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
4861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
4921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
4981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
5761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
5881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
5941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
6001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
6061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
6121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
6181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
6241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
6301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
6361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
6421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
6481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
6541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
6601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
6661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
6721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
6781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
6841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
6901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
6961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
7021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
7081 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
7141 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
7201 tagtgcgcga gcaaaattta agcta
//
