| 质粒名称: | pENN.AAV.cTNT.PI.eGFP.WPRE.rBG |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 105543 |
| 存储实验室: | James M. Wilson |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,AAV |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | cTNT-PI-EGFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | cTNT |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pENN.AAV.cTNT.PI.eGFP.WPRE.rBG was a gift from James M. Wilson (Addgene plasmid # 105543 ; http://n2t.net/addgene:105543 ; RRID:Addgene_105543) |
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.cTNT.PI.eGFP.WPRE.rBG 5573 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION AAV expression of EGFP from cTNT promoter.
ACCESSION .
VERSION .
KEYWORDS pENN.AAV.cTNT.PI.eGFP.WPRE.rBG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5573)
TITLE AAV plasmids
REFERENCE 2 (bases 1 to 5573)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5573
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 113..134
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 149..179
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 187..203
/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 192..214
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 211..227
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 211..227
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
repeat_region 265..394
/label=AAV2 ITR
intron 1036..1168
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 1151..1170
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1213..1231
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 1254..1263
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1260..1979
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1305..1326)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1566..1585)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1913..1934
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2060..2648
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2113..2133)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2531..2542)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2674..2693)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2739..2794
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
repeat_region 2887..3016
/label=AAV2 ITR
primer_bind complement(3035..3052)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3035..3051)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3044..3066)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3193..3648
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3280..3299)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3490..3511
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3674..3778
/gene="bla"
/label=AmpR promoter
CDS 3779..4639
/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(3997..4016)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4810..5398
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5299..5318
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5552..5569
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
241 gatttaatta aggccttaat taggctgcgc gctcgctcgc tcactgaggc cgcccgggca
301 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
361 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
421 tacttatcta ccagggtaat ggggatcctc tagaactata gctagaattc gcccttacgg
481 gccccccctc gaggtcggga taaaagcagt ctgggctttc acatgacagc atctggggct
541 gcggcagagg gtcgggtccg aagcgctgcc ttatcagcgt ccccagccct gggaggtgac
601 agctggctgg cttgtgtcag cccctcgggc actcacgtat ctccgtccga cgggtttaaa
661 atagcaaaac tctgaggcca cacaatagct tgggcttata tgggctcctg tgggggaagg
721 gggagcacgg agggggccgg ggccgctgct gccaaaatag cagctcacaa gtgttgcatt
781 cctctctggg cgccgggcac attcctgctg gctctgcccg ccccggggtg ggcgccgggg
841 ggaccttaaa gcctctgccc cccaaggagc ccttcccaga cagccgccgg cacccaccgc
901 tccgtgggac gatccccgaa gctctagagc tttattgcgg tagtttatca cagttaaatt
961 gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
1021 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
1081 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
1141 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
1201 ggctagagta cttaatacga ctcactatag gctagcctcg agaattcacg cgggccgcca
1261 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
1321 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
1381 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
1441 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
1501 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
1561 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
1621 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
1681 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
1741 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
1801 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
1861 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
1921 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
1981 aagctcgcgt ggtacctcta gagtcgaccc gggcggcctc gaggacgggg tgaactacgc
2041 ctgaatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
2101 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
2161 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
2221 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
2281 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
2341 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
2401 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
2461 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
2521 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
2581 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
2641 ctccccgcat cgataccgtc gatgaggatc cgatcttttt ccctctgcca aaaattatgg
2701 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
2761 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga agcaattcgt tgatctgaat
2821 ttcgaccacc cataataccc attaccctgg tagataagta gcatggcggg ttaatcatta
2881 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
2941 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
3001 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
3061 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
3121 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
3181 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
3241 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
3301 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
3361 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
3421 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
3481 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
3541 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
3601 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
5461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
5521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aag
//
