| 质粒名称: | pSR660 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 65216 |
| 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（未销毁） |
| 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: | pSR660 was a gift from Robert Kotin (Addgene plasmid # 65216 ; http://n2t.net/addgene:65216 ; RRID:Addgene_65216) |
| 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 pSR660 9362 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2
rep gene and polycistronic AAV-8 cap gene in Sf9 insect cells.
ACCESSION .
VERSION .
KEYWORDS pSR660
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9362)
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 9362)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9362
/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(6627..6736)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 6760..6851
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 6776..6793
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
polyA_signal 8952..9086
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8989..9008)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9043..9062
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 9115..9280
/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 agccgcagag accaaagttc
4321 aactgaaacg aatcaaccgg tttattgatt aacaggcaat tacagattac gggtgaggta
4381 acgggtgcca atggggcggg gttcagagta cacgccttct gtattaacag caaagtccac
4441 acttgtagat ttgtagtagt tggaggtgta ctggatctcg gggttccagc gcttgctgtt
4501 ttccttctgc agctcccatt caatttccac gctgacctgt ccggtgctgt attgcgtgat
4561 gaaagagttc agctttgact ggttgaaggt ggtcggagga tccgcaggta caggcgtgtt
4621 cttgatcagg atctgaggcg gaggatgttt caggccaaag ccgcccatca gcggagacgg
4681 gtggaagttg ccgtccgtgt gaggaatctt ggcccagatg ggaccctgca ggtacacgtc
4741 ccggttctgc cagaccatac cgggtaaggc cccctggctg ttgacagttc caatttgagg
4801 agccgtgttt tgctgctgca agttatctgc cacgataccg tattcctctg tagccacagg
4861 gttagtggtt ttgatttctt cctcgctggt gagcatgaca tcgctgtaat ccgcattgtc
4921 tctggcagca ttttgtttgc caaaaatcag gatcccgtta ctgggaaaaa aacgctcctc
4981 gtcgtctttg tgtgttgcca tagcgatgcc aggattagcc aatgaatttc ttccattcag
5041 atggtatttg gtcccagcag tccaggcaaa gttgctattg ttgttttgcc cggttgtcgt
5101 tgagacgcgt tgttggcggt aacagggtcc tggcagccag ttctttgcct gattggccat
5161 tgtattaggc ccaccttggc tgaagcccag agtctgcgta tttgccgtgc ctcctgttgt
5221 ttgagtccga gacaagtagt acaggtactg gtcaatcaga ggattcatca gccggtccaa
5281 gctctggctg tgggcgtagc tgctgtggaa aggcacgtcc tcgaaggtgt aagtaaactg
5341 gaagttgttg ccggttctca gcatctgcga aggaaagtat tccaggcagt agaaggagga
5401 gcgtcccacg gcctgactac cgttgttgag tgttaggtag ccgtactggg gaatcatgaa
5461 cacgtccgcc gggaacggag gcaggcagcc ctggtgggca gagccgagaa cgtacggcag
5521 ctggtactcc gagtccgtaa acacctggat ggtgctggtg aggttattgg cgatggtctt
5581 ggtgccttca ttctgcgtga cctccttgac ctggatgttg aagagcttga agctgagtct
5641 cttgggccgg aatccccagt tgttgttgat gagtcgctgc cagtcacgtg gtgaaaagtg
5701 gcagtggaat ctgttaaagt caaaataccc ccagggggtg ctgtagccga agtaggtgtt
5761 gtcgttggtg gctcctcccg atgtcccgtt ggagatttgc ttgtagaggt ggttgttgta
5821 ggtgggcagg gcccaggttc gggtgctggt ggtgatgact ctgtcgccca gccatgtgga
5881 atcgcaatgc caatttcccg aggaactacc cactccgtcg gcgccttcgt tattgtctgc
5941 cattggtgcg ccaccgcctg cagccattgt attaggtccc acaccagagg gcgctgctgg
6001 aggttctccg agaggttgag ggtctggaac tgactctgag tcgccagtct gaccaaaatt
6061 gagtcttttt ctggcgggct gttggccttt cttgccgatg cccgtagagg agtctggaga
6121 acgctggggt gatggctcta ccggtctctt ctttccagga gccgtcttag cgccttcctc
6181 aaccagaccg agaggttcga gaacccgctt cttggcctgg aagactgctc gcccgaggtt
6241 gcccccaaaa gacgtatctt cttgcagacg ctcctggaat tcagcgtcgg cgtggttata
6301 acgaaggtaa gggttgtcgc cggcctgcag ttgctggtcg taagccttat cgtgctcaag
6361 ggcagcggcg tccgcagcgt tcacgggttc gcccttgtcc agcccgttga agggtcctag
6421 gtatttgtaa ccggggagaa ccaggccgcg tccgtcgtct tgtttctgtt ggttagcctt
6481 tggcttagga gcgccgggtt tgagggccca ccactcgcga atgccctcag agaggttgtc
6541 ctcgagccaa tcgggtagat aaccgtcggc agccgtggcg gcaggaatcc cgcgcgcttc
6601 cccgggtgat caagtcttcg tcgagtgatt gtaaataaaa tgtaatttac agtatagtat
6661 tttaattaat atacaaatga tttgataata attcttattt aactataata tattgtgttg
6721 ggttgaatta aaggtccgta tactccggaa tattaataga tcatggagat aattaaaatg
6781 ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa
6841 aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga tctgccgccc
6901 tggcggggtt ttacgagatt gtgattaagg tccccagcga ccttgacgag catctgcccg
6961 gcatttctga cagctttgtg aactgggtgg ccgagaagga gtgggagttg ccgccagatt
7021 ctgacttgga tctgaatctg attgagcagg cacccctgac cgtggccgag aagctgcagc
7081 gcgactttct gacggagtgg cgccgtgtga gtaaggcccc ggaggccctt ttctttgtgc
7141 aatttgagaa gggagagagc tacttccact tacacgtgct cgtggaaacc accggggtga
7201 aatccttagt tttgggacgt ttcctgagtc agattcgcga aaaactgatt cagagaattt
7261 accgcgggat cgagccgact ttgccaaact ggttcgcggt cacaaagacc agaaacggcg
7321 ccggaggcgg gaacaaggtg gtggacgagt gctacatccc caattacttg ctccccaaaa
7381 cccagcctga gctccagtgg gcgtggacta atttagaaca gtatttaagc gcctgtttga
7441 atctcacgga gcgtaaacgg ttggtggcgc agcatctgac gcacgtgtcg cagacgcagg
7501 agcagaacaa agagaatcag aatcccaatt ctgacgcgcc ggtgatcaga tcaaaaactt
7561 cagccaggta catggagctg gtcgggtggc tcgtggacaa ggggattacc tcggagaagc
7621 agtggatcca ggaggaccag gcctcataca tctccttcaa tgcggcctcc aactcgcggt
7681 cccaaatcaa ggctgccttg gacaatgcgg gaaagattat gagcctgact aaaaccgccc
7741 ccgactacct ggtgggccag cagcccgtgg aggacatttc cagcaatcgg atttataaaa
7801 ttttggaact aaacgggtac gatccccaat atgcggcttc cgtctttctg ggatgggcca
7861 cgaaaaagtt cggcaagagg aacaccatct ggctgtttgg gcctgcaact accgggaaga
7921 ccaacatcgc ggaggccata gcccacactg tgcccttcta cgggtgcgta aactggacca
7981 atgagaactt tcccttcaac gactgtgtcg acaagatggt gatctggtgg gaggagggga
8041 agatgaccgc caaggtcgtg gagtcggcca aagccattct cggaggaagc aaggtgcgcg
8101 tggaccagaa atgcaagtcc tcggcccaga tagacccgac tcccgtgatc gtcacctcca
8161 acaccaacat gtgcgccgtg attgacggga actcaacgac cttcgaacac cagcagccgt
8221 tgcaagaccg gatgttcaaa tttgaactca cccgccgtct ggatcatgac tttgggaagg
8281 tcaccaagca ggaagtcaaa gactttttcc ggtgggcaaa ggatcacgtg gttgaggtgg
8341 agcatgaatt ctacgtcaaa aagggtggag ccaagaaaag acccgccccc agtgacgcag
8401 atataagtga gcccaaacgg gtgcgcgagt cagttgcgca gccatcgacg tcagacgcgg
8461 aagcttcgat caactacgca gacaggtacc aaaacaaatg ttctcgtcac gtgggcatga
8521 atctgatgct gtttccctgc agacaatgcg agagaatgaa tcagaattca aatatctgct
8581 tcactcacgg acagaaagac tgtttagagt gctttcccgt gtcagaatct caacccgttt
8641 ctgtcgtcaa aaaggcgtat cagaaactgt gctacattca tcatatcatg ggaaaggtgc
8701 cagacgcttg cactgcctgc gatctggtca atgtggattt ggatgactgc atctttgaac
8761 aataaactcg aggaatcaag cttgcatgcc tgcaggtcga ctctagagcc tgcagtctcg
8821 acaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
8881 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
8941 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
9001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
9061 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
9121 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
9181 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
9241 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
9301 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
9361 tt
//
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