| 质粒名称: | pENN.AAV.CB7.CI.mCherry.WPRE.RBG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 105544 |
| 存储实验室: | James M. Wilson |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 完整质粒大小: | 6502bp |
| 载体类型: | 哺乳细胞表达、 AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | CB7-CI-mCherry |
| 启动子: | CB7 |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: |
pENN.AAV.CB7.CI.mCherry.WPRE.RBG was a gift from James M. Wilson (Addgene plasmid # 105544 ; http://n2t.net/addgene:105544 ; RRID:Addgene_105544)
For viral preps, please replace (Addgene plasmid # 105544) in the above sentence with: (Addgene viral prep # 105544-AAV1) or (Addgene viral prep # 105544-AAV9)
|
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 pENN.AAV.CB7.CI.mCherry.WPRE.RBG 6502 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV expression of mCherry from CB7 promoter.
ACCESSION .
VERSION .
KEYWORDS pENN.AAV.CB7.CI.mCherry.WPRE.RBG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6502)
TITLE AAV plasmids
REFERENCE 2 (bases 1 to 6502)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6502
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR
enhancer 204..583
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 585..862
/label=chicken beta-actin promoter
intron 863..1879
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1803..1825
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1887..1906
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1962..2672
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2107..2125)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2375..2394
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 2725..3313
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2778..2798)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3196..3207)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3339..3358)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3404..3459
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
repeat_region 3552..3681
/label=AAV2 ITR
primer_bind complement(3700..3717)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3700..3716)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3709..3731)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3858..4313
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3945..3964)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4155..4176
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4339..4443
/gene="bla"
/label=AmpR promoter
CDS 4444..5304
/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 complement(4662..4681)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5475..6063
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5964..5983
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6217..6234
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6351..6372
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6387..6417
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6425..6441
/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)."
primer_bind 6430..6452
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6449..6465
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6449..6465
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
181 atcctctaga actatagcta gtcgacattg attattgact agttattaat agtaatcaat
241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
541 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
601 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
661 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
721 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
781 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
841 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
901 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
961 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
1021 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
1081 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
1141 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
1201 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
1261 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
1321 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
1381 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
1441 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
1501 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
1561 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
1621 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
1681 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
1741 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
1801 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
1861 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
1921 tggcaaagaa ttcacgctag atttaagaag gagatataca tatggtgagc aagggcgagg
1981 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
2041 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
2101 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
2161 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
2221 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
2281 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
2341 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
2401 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
2461 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
2521 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
2581 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
2641 ccaccggcgg catggacgag ctgtacaagt aagcatgcaa gcttcgagga cggggtgaac
2701 tacgcctgaa tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
2761 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
2821 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
2881 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
2941 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
3001 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
3061 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
3121 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
3181 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
3241 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
3301 gccgcctccc cgcatcgata ccgtcgatga ggatccgatc tttttccctc tgccaaaaat
3361 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
3421 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaagcaa ttcgttgatc
3481 tgaatttcga ccacccataa tacccattac cctggtagat aagtagcatg gcgggttaat
3541 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
3601 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
3661 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
3721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
3781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
3841 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
3901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
3961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
4021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
4081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
4141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
4201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
4261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
4321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
4381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
4441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
4501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
4561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
4621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
4681 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
4741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
4801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
4861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
4921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
4981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
5041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
5101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
5161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
5221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
5281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
6181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
6241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
6301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
6361 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
6421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
6481 tttaattaag gccttaatta gg
//
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