| 质粒名称: | pSR646 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 65215 |
| 存储实验室: | Robert Kotin |
|
载体骨架基本信息 |
|
| 载体名称: | pFastBac-Dual |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5238 bp |
| 完整质粒大小: | 9335 bp |
| 载体修饰: | -- |
| 载体类型: | 昆虫表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | modified AAV-2 rep78 |
| 物种来源: | -- |
| 插入大小: | 1866 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | polyhedrin |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI（在克隆过程中销毁） |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | AAATGATAACCATCTCGC |
| 3' 测序引物 | GAAATTTGTGATGCTATTGC |
|
文献引用方法 |
|
| 材料和方法部分: | pSR646 was a gift from Robert Kotin (Addgene plasmid # 65215 ; http://n2t.net/addgene:65215 ; RRID:Addgene_65215) |
| 参考文献部分: | A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells. Smith RH, Levy JR, Kotin RM. Mol Ther. 2009 Nov;17(11):1888-96. doi: 10.1038/mt.2009.128. Epub 2009 Jun 16. 10.1038/mt.2009.128 PubMed 19532142 |
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 pSR646 9284 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2
rep gene and polycistronic AAV-6 cap gene in Sf9 insect cells.
ACCESSION .
VERSION .
KEYWORDS pSR646
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9284)
AUTHORS Smith RH, Levy JR, Kotin RM
TITLE A simplified baculovirus-AAV expression vector system coupled with
one-step affinity purification yields high-titer rAAV stocks from
insect cells.
JOURNAL Mol Ther. 2009 Nov;17(11):1888-96. doi: 10.1038/mt.2009.128. Epub
2009 Jun 16.
PUBMED 19532142
REFERENCE 2 (bases 1 to 9284)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9284
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 102..557
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(189..208)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 399..420
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 584..688
/gene="bla"
/label=AmpR promoter
CDS 689..1549
/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(907..926)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1720..2308
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2209..2228
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2462..2479
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 2611..2835
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2902..3435)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(3624..3652)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal 4161..4209
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4234..4253
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
promoter complement(6549..6658)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 6682..6773
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 6698..6715
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
polyA_signal 8874..9008
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8911..8930)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8965..8984
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 9037..9202
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2521 tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg
2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
3121 ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc
3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc
3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac
4021 gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga
4081 aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg
4141 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
4201 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
4261 tcccccatct cccggtaccg catgctatgc atcagctgct agcttacagg ggacgggtga
4321 ggtaacgggt gccaatgggg cgaggctcag tataaagtcc attgttgtcc acagtgaaat
4381 caacgttggc agattttgca tagttagatg tatactgcac ttcgggattc cagcgtttgc
4441 tgttttcttt ctgcagctcc cattcaatct ccacgctcac ttgtcctgtg gaatactggg
4501 tgatgaatga agcaaacttt gtagccgaaa actctgccgg aggattcgca ggaacaggcg
4561 tgtttttgat gaggatctga ggaggcgggt gcttaagtcc aaagccgccc atgagaggag
4621 acgggtgaaa gtgtccatcc gtgtgaggaa ttttggccca aataggaccc tgcaggtata
4681 cgtctctgtc ttgccacacc attccaggta aggctcccat aacatgcaca tctccggtcg
4741 cagggtctgt gctgctgctc tggagattga ctgccacagt cccaaatctt tcggtggcca
4801 cggggttagt ggctttgatt tcctcttcgt ctgtgatcat gacattgtcc aatgcagtgt
4861 ttgaagctcc ggcgctctcc tttccaaaaa tcatgacacc gctcatggga aagaacttgt
4921 ctttgtcgtc tttgtgtgag gccatagcag tgccagggtt gattatagat tcacgcccat
4981 taaggttata ttttgaagca ccagtccagg taaagttgct gttgttgttg tctgtttttg
5041 ttttagaaac gcgctgctgc cggtaacagg gtccaggtag ccagtttttg ggctgaacag
5101 acatgccagc tggagacccc cggctaaaca gcaagtcctt gttttgggca cttccggact
5161 gattctgagt tctgttcagg taatacaggt actggtcgat gagaggattc atcagccggt
5221 ccaggctctg gctgtgcgcg tagctgctgt ggaaaggcac gtcctcgaag gtgtagctga
5281 aggtaaagtt attgcccgtt ctcagcatct gcgatgggaa atattccagg cagtaaaagg
5341 atgaccgtcc cactgcctgg ctgccattgt tgagcgttag gtagccgtac tgcggaatca
5401 tgaacacgtc cgccgggaac ggagggaggc agccctggtg cgcagagccg aggacgtacg
5461 gcaactggta ctccgagtcc gagaagactt gaaccgtgct ggtaaggtta ttagcgatgg
5521 tcgtgacgcc atcattcgtc gtgacctcct tgacttggat gttgaagagc ttgaagttga
5581 gtctcttggg ccggaatccc caattgttgt tgatgagtcg ctgccagtca cgtggtgaga
5641 aatggcagtg gaatctgttg aaatcaaaat acccccaggg ggtgctgtag ccgaagtagt
5701 ggttgtcgtt gctggccccc gttgaagcac tggagatttg cttgtagagg tggttgttat
5761 aggtgggcaa ggcccatgtt cgggtgctgg tggtgatgac tctgtcgccc agccatgtgg
5821 aatcgcaatg ccaatttcct gaggcattac ccactccgtc ggcgccttcg ttattgtctg
5881 ccattggtgc gccaccgcct gaagccattg tagtaggtcc cacagcagcg ggggttgctg
5941 gaggttctcc gagaggttgt gggtcgggga ctgactctga gtcgccagtc tgaccaaaat
6001 tgagtctctt tttagcgggc tgctggcctg tcttgccaat gcccgaggag gagtctggct
6061 cttgtggcga ctgctctacc ggacgtttct ttccaggagc cgtcttagca ccttcctcaa
6121 ccagaccaaa aggttcgaga accctcttct tggcctggaa gactgctcgc ccgaggttgc
6181 ccccaaaaga cgtatcttct tgcagacgct cctgaaactc ggcgtcggcg tggttatacc
6241 gcaggtacgg attgtcaccc gctttgagct gctggtcgta ggccttgtcg tgctcgaggg
6301 ccgctgcatc cgccgcgttg acgggctccc ccttgtcgag tccgttgaag ggtccgaggt
6361 acttgtagcc aggaagcacc agaccccggc cgtcgtcctg cttttgctgg ttggctttgg
6421 gtttcggggc tccaggtttg aggtcccacc actcgcgaat gccctcagag aggttgtcct
6481 cgagccaatc gggtagataa ccgtcggcag ccaaggcggc ggcccgggtg atcaagtctt
6541 cgtcgagtga ttgtaaataa aatgtaattt acagtatagt attttaatta atatacaaat
6601 gatttgataa taattcttat ttaactataa tatattgtgt tgggttgaat taaaggtccg
6661 tatactccgg aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata
6721 aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg
6781 attattcata ccgtcccacc atcgggcgcg gatctgccgc cctggcgggg ttttacgaga
6841 ttgtgattaa ggtccccagc gaccttgacg agcatctgcc cggcatttct gacagctttg
6901 tgaactgggt ggccgagaag gagtgggagt tgccgccaga ttctgacttg gatctgaatc
6961 tgattgagca ggcacccctg accgtggccg agaagctgca gcgcgacttt ctgacggagt
7021 ggcgccgtgt gagtaaggcc ccggaggccc ttttctttgt gcaatttgag aagggagaga
7081 gctacttcca cttacacgtg ctcgtggaaa ccaccggggt gaaatcctta gttttgggac
7141 gtttcctgag tcagattcgc gaaaaactga ttcagagaat ttaccgcggg atcgagccga
7201 ctttgccaaa ctggttcgcg gtcacaaaga ccagaaacgg cgccggaggc gggaacaagg
7261 tggtggacga gtgctacatc cccaattact tgctccccaa aacccagcct gagctccagt
7321 gggcgtggac taatttagaa cagtatttaa gcgcctgttt gaatctcacg gagcgtaaac
7381 ggttggtggc gcagcatctg acgcacgtgt cgcagacgca ggagcagaac aaagagaatc
7441 agaatcccaa ttctgacgcg ccggtgatca gatcaaaaac ttcagccagg tacatggagc
7501 tggtcgggtg gctcgtggac aaggggatta cctcggagaa gcagtggatc caggaggacc
7561 aggcctcata catctccttc aatgcggcct ccaactcgcg gtcccaaatc aaggctgcct
7621 tggacaatgc gggaaagatt atgagcctga ctaaaaccgc ccccgactac ctggtgggcc
7681 agcagcccgt ggaggacatt tccagcaatc ggatttataa aattttggaa ctaaacgggt
7741 acgatcccca atatgcggct tccgtctttc tgggatgggc cacgaaaaag ttcggcaaga
7801 ggaacaccat ctggctgttt gggcctgcaa ctaccgggaa gaccaacatc gcggaggcca
7861 tagcccacac tgtgcccttc tacgggtgcg taaactggac caatgagaac tttcccttca
7921 acgactgtgt cgacaagatg gtgatctggt gggaggaggg gaagatgacc gccaaggtcg
7981 tggagtcggc caaagccatt ctcggaggaa gcaaggtgcg cgtggaccag aaatgcaagt
8041 cctcggccca gatagacccg actcccgtga tcgtcacctc caacaccaac atgtgcgccg
8101 tgattgacgg gaactcaacg accttcgaac accagcagcc gttgcaagac cggatgttca
8161 aatttgaact cacccgccgt ctggatcatg actttgggaa ggtcaccaag caggaagtca
8221 aagacttttt ccggtgggca aaggatcacg tggttgaggt ggagcatgaa ttctacgtca
8281 aaaagggtgg agccaagaaa agacccgccc ccagtgacgc agatataagt gagcccaaac
8341 gggtgcgcga gtcagttgcg cagccatcga cgtcagacgc ggaagcttcg atcaactacg
8401 cagacaggta ccaaaacaaa tgttctcgtc acgtgggcat gaatctgatg ctgtttccct
8461 gcagacaatg cgagagaatg aatcagaatt caaatatctg cttcactcac ggacagaaag
8521 actgtttaga gtgctttccc gtgtcagaat ctcaacccgt ttctgtcgtc aaaaaggcgt
8581 atcagaaact gtgctacatt catcatatca tgggaaaggt gccagacgct tgcactgcct
8641 gcgatctggt caatgtggat ttggatgact gcatctttga acaataaact cgaggaatca
8701 agcttgcatg cctgcaggtc gactctagag cctgcagtct cgacaagctt gtcgagaagt
8761 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
8821 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
8881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
8941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
9001 tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga aatctagttc
9061 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
9121 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
9181 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
9241 atcctgccaa ctccatgtga caaaccgtca tcttcggcta cttt
//
