| 质粒名称: | rEstus |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 209767 |
| 存储实验室: | Stefan Heinemann |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(+)_Puro |
| 完整质粒大小: | 6427 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | rEstus / ASAP3-N138G-Y141T-S318A-Q396R |
| 物种来源: | G. gallus (chicken); A.victoria |
| 插入大小: | 1281 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | NotI(未破坏) |
| 5’测序引物 | GGC TAA CTA GAG AAC CCA CTG |
| 3’测序引物 | TAG AAG GCA CAG TCG AGG |
|
文献引用方法 |
|
| 材料和方法部分: | rEstus was a gift from Stefan Heinemann (Addgene plasmid # 209767 ; http://n2t.net/addgene:209767 ; RRID:Addgene_209767) |
| 参考文献部分: | An ultrasensitive voltage indicator uncovers the electrical activity of non-excitable cells. Rühl P., Nair, A. G., Gawande, N., Dehiwalage, S. N. C. W., Münster, L., Schönherr, R., & Heinemann, S. H.. bioRxiv 2023. 10.1101/2023.10.05.560122 |
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 rEstus 6427 bp ds-DNA circular SYN 06-NOV-2023
DEFINITION To express a bright and sensitive genetically encoded voltage
indicator in mammalian cells..
ACCESSION .
VERSION .
KEYWORDS rEstus
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6427)
TITLE An ultrasensitive voltage indicator uncovers the electrical activity
of non-excitable cells
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6427)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 6, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6427
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2283..2300)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2289..2513
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2559..2987
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2646..2665)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2856..2877
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2996..3016)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3001..3330
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3181..3316
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3243..3262
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3341..3940
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3341..3360)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3837..3857
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4100..4221
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4137..4156)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4191..4210
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4270..4286)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4270..4286)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4283..4305)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4294..4310
/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(4318..4348)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4363..4384
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4501..4518)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4672..5260)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4752..4771)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5431..6291)
/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 6054..6073
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6292..6396)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcatg
961 gagacgactg tgaggtatga acaggggtca gagctcacta aaacttcgag ctctccaaca
1021 gcagatgagc ccacgataaa gattgatgat ggtcgtgatg agggtaatga acaagacagc
1081 tgttccaata ccattaggag aaaaatttcc ccgtttgtga tgtcatttgg attcagagta
1141 tttggagttg tgcttattat tgtagacatc atagtggtga ttgtggatct ggccatcagt
1201 gagaagaaaa gaggcattag agagattctt gaaggtgttt ccctggctat agcactcttc
1261 ttccttgttg atgttctcat gagagtgttt gttgaaggct tcaagaacta tttccggtcc
1321 aaactgaata ctttggatgc agtcatagta gtgggcactc tgctaattgg tatgaccacc
1381 tccttctctg acggtacttt tcgcggcgat aacgtgtata ttaccgcgga taaacagaaa
1441 aacggcatta aagcgaactt taccgtgcgc cataacgtgg aagatggcag cgtgcagctg
1501 gcggatcatt atcagcagaa caccccgatt ggcgatggcc cggtgctgct accggataac
1561 cattatctga gcacccagac cgtgctgagc aaagatccga acgaaaaacg cgatcacatg
1621 gtgctgctgg aatttgtgac cgcagcgggc attacacacg gcatggatga actgtatggc
1681 ggcaccggcg gcagcgcgag ccagggcgaa gaactgttta ccggcgtggt gccgattctg
1741 gtggaactgg atggcgatgt gaacggccat aaatttagcg tgcgcggcga aggcgaaggc
1801 gatgcgacca ttggcaaact gaccctgaaa tttatttgca ccaccggcaa actaccggtg
1861 ccgtggccga ccctggtgac caccttaacc tatggcgtgc agtgctttgc ccgctatccg
1921 gatcatatga aacgccatga tttttttaaa agcgcgatgc cggaaggcta tgtgcaggaa
1981 cgcaccatta gctttaaaga tgatggcaaa tataaaaccc gcgcggtggt gaaatttgaa
2041 ggcgataccc tggtgaaccg cattgaactg aaaggcaccg attttaaaga agatggcaac
2101 attctggggc ataaactgga atataacaca gatcagatgc cgcggatggt tactcttctt
2161 cgagttctga gaattgttat cttaataaga atatttcagc tggcttcaca gaagaaacaa
2221 cttgaagtgg taacctaagc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
2281 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
2341 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
2401 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
2461 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
2521 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
2581 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
2641 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
2701 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
2761 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
2821 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
2881 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
2941 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
3001 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
3061 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
3121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
3181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
3241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
3301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgc atgaccgagt acaagcccac
3361 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
3421 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
3481 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
3541 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
3601 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
3661 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
3721 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
3781 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
3841 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
3901 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc
3961 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
4021 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
4081 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
4141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
4201 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
4261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
4321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
4381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
4441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
4501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
5101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
5221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
5281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
5341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
5401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
5461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
5521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
5581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
5641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
5701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
5761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
5821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
5881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
5941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
6001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
6061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
6121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
6181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
6241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
6301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
6361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
6421 tgacgtc
//
