| 质粒名称: | pcDNA3.1/Puro-CAG-ASAP1 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 52519 |
| 存储实验室: | Michael Lin |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1/Puro-CAG |
| 载体出品公司: | Invitrogen/Dr. Paulmurugan Ramasamy |
| 空载体大小: | 6107 bp |
| 完整质粒大小: | 7380 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: | XL10 Gold |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Accelerated Sensor of Action Potentials 1 / ASAP1 |
| 物种来源: | Synthetic; Aequorea victoria |
| 插入大小: | 1281 bp |
| 启动子: | CAG |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI (未破坏) |
| 3’克隆位点: | HindIII(未破坏) |
| 5’测序引物 | AAGGTGGTGGCTGGTGTGGC |
| 3’测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1/Puro-CAG-ASAP1 was a gift from Michael Lin (Addgene plasmid # 52519 ; http://n2t.net/addgene:52519 ; RRID:Addgene_52519) |
| 参考文献部分: | High-fidelity optical reporting of neuronal electrical activity with an ultrafast fluorescent voltage sensor. St-Pierre F, Marshall JD, Yang Y, Gong Y, Schnitzer MJ, Lin MZ. Nat Neurosci. 2014 Apr 22. doi: 10.1038/nn.3709. 10.1038/nn.3709 PubMed 24755780 |
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 pcDNA3.1/Puro-CAG-ASAP1 7380 bp ds-DNA circular SYN 28-JUL-2023
DEFINITION For mammalian expression of the fluorescent voltage sensor ASAP1.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1/Puro-CAG-ASAP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7380)
AUTHORS St-Pierre F, Marshall JD, Yang Y, Gong Y, Schnitzer MJ, Lin MZ
TITLE High-fidelity optical reporting of neuronal electrical activity with
an ultrafast fluorescent voltage sensor.
JOURNAL Nat Neurosci. 2014 Apr 22. doi: 10.1038/nn.3709.
PUBMED 24755780
REFERENCE 2 (bases 1 to 7380)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 28, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7380
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 8..17
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1412..1429)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1418..1642
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1688..2116
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1775..1794)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1985..2006
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2125..2145)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2130..2459
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2310..2445
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2372..2391
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2470..3069
/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(2470..2489)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2966..2986
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 3229..3350
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3266..3285)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3320..3339
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3399..3415)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3399..3415)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3412..3434)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3423..3439
/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(3447..3477)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3492..3513
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3630..3647)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3801..4389)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3881..3900)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4560..5420)
/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 5183..5202
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5421..5525)
/gene="bla"
/label=AmpR promoter
enhancer 5557..5936
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 6212..7228
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 7152..7174
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 7236..7255
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
ORIGIN
1 gctagccgcc accatggaga cgactgtgag gtatgaacag gggtcagagc tcactaaaac
61 ttcgagctct ccaacagcag atgagcccac gataaagatt gatgatggtc gtgatgaggg
121 taatgaacaa gacagctgtt ccaataccat taggagaaaa atttccccgt ttgtgatgtc
181 atttggattc agagtatttg gagttgtgct tatcattgta gacatcatag tggtgattgt
241 ggatctggcc atcagtgaga agaaaagagg cattagagag attcttgaag gtgtttccct
301 ggctatagca ctcttcttcc ttgttgatgt tctcatgaga gtgtttgttg aaggcttcaa
361 gaactatttc cggtccaaac tgaatacttt ggatgcagtc atagtagtgg gcactctgct
421 aattaatatg acctactcct tctctgacct tgctgccttt aacagccata acgtgtatat
481 taccgcggat aaacagaaaa acggcattaa agcgaacttt accgtgcgcc ataacgtgga
541 agatggcagc gtgcagctgg cggatcatta tcagcagaac accccgattg gcgatggccc
601 ggtgctgctg ccggataacc attatctgag cacccagacc gtgctgagca aagatccgaa
661 cgaaaaacgc gatcacatgg tgctgctgga atttgtgacc gcagcgggca ttacacacgg
721 catggatgaa ctgtatggcg gcaccggcgg cagcgcgagc cagggcgaag aactgtttac
781 cggcgtggtg ccgattctgg tggaactgga tggcgatgtg aacggccata aatttagcgt
841 gcgcggcgaa ggcgaaggcg atgcgaccat tggcaaactg accctgaaat ttatttgcac
901 caccggcaaa ctaccggtgc cgtggccgac cctggtgacc accttaacct atggcgtgca
961 gtgctttagc cgctatccgg atcatatgaa acgccatgat ttttttaaaa gcgcgatgcc
1021 ggaaggctat gtgcaggaac gcaccattag ctttaaagat gatggcaaat ataaaacccg
1081 cgcggtggtg aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggcaccga
1141 ttttaaagaa gatggcaaca ttctggggca taaactggaa tataacacag atcagatgcc
1201 gcagatggtt actcttcttc gagttctgag aattgttatc ttaataagaa tatttcgcct
1261 ggcttcacag aagaaacaac ttgaagtggt aacctaatga aagcttggta ccgagctcgg
1321 atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag
1381 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
1441 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
1501 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
1561 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
1621 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
1681 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
1741 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
1801 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
1861 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
1921 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
1981 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
2041 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
2101 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
2161 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
2221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2281 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
2341 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
2401 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
2461 agctcccgca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
2521 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
2581 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
2641 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
2701 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
2761 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
2821 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
2881 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
2941 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
3001 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
3061 ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
3121 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
3181 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
3241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
3301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
3361 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
3421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
3481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
3541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
3601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
3661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
3721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
3781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
3841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
3901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
3961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
4261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
4501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
4561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
4621 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
4681 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
4741 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
4801 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
4861 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
4921 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
4981 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
5041 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
5101 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
5161 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5221 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5281 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5341 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
5401 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
5461 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
5521 ccgcgcacat ttccccgaaa agtgccacct gacgtcgaca ttgattattg actagttatt
5581 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
5641 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
5701 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
5761 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
5821 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
5881 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
5941 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
6001 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggsgc
6061 gccrggsggg gggggggggg ggggggggsg gggsggggsg aggsggagag gtgsggcggc
6121 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
6181 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
6241 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
6301 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
6361 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
6421 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
6481 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
6541 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
6601 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
6661 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
6721 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
6781 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg rgggctcggg
6841 ggrggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
6901 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
6961 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
7021 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
7081 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
7141 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
7201 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
7261 tcatcatttt ggcaaagaat tcctcgagga attcactcct caggtgcagg ctgcctatca
7321 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctgcatga
//
