| 质粒名称: | pSR657 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 65214 |
| 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(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: | pSR657 was a gift from Robert Kotin (Addgene plasmid # 65214 ; http://n2t.net/addgene:65214 ; RRID:Addgene_65214) |
| 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 pSR657 9286 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2
rep gene and polycistronic AAV-2 cap gene in Sf9 insect cells.
ACCESSION .
VERSION .
KEYWORDS pSR657
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9286)
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 9286)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9286
/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(6551..6660)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 6684..6775
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 6700..6717
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
polyA_signal 8876..9010
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8913..8932)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8967..8986
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 9039..9204
/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 agcttacaga ttacgagtca
4321 ggtatctggt gccaatgggg cgaggctctg aatacacgcc attagtgtcc acagtaaagt
4381 ccacattaac agacttgttg tagttggaag tgtactgaat ttcgggattc cagcgtttgc
4441 tgttttcctt ctgcagctcc cactcgatct ccacgctgac ctgtcccgtg gagtactgtg
4501 tgatgaagga agcaaacttt gccgcactga aggtggtcga aggattcgca ggtaccgggg
4561 tgttcttgat gagaatctgt ggaggagggt gtttaagtcc gaatccaccc atgaggggag
4621 aggggtgaaa atgtccgtcc gtgtgtggaa tctttgccca gatgggcccc tgaaggtaca
4681 catctctgtc ctgccagacc atgcctggaa gaacgccttg tgtgttgaca tctgcggtag
4741 ctgcttgtct gttgcctctc tggaggttgg tagatacaga accatactgc tccgtagcca
4801 cgggattggt tgtcctgatt tcctcttcgt ctgtaatcat gaccttttca atgtccacat
4861 ttgttttctc tgagccttgc ttcccaaaga tgagaacccc gctctgagga aaaaactttt
4921 cttcatcgtc cttgtggctt gccatggccg ggcccggatt caccagagag tctctgccat
4981 tgaggtggta cttggtagct ccagtccacg agtattcact gttgttgtta tccgcagatg
5041 tctttgatac tcgctgctgg cggtaacagg gtccaggaag ccagttccta gactggtccc
5101 gaatgtcact cgctccggcc tgagaaaact gaagccttga ctgcgtggtg gttccacttg
5161 gagtgtttgt tctgctcaag taatacaggt actggtcgat gagaggattc atgagacggt
5221 ccagactctg gctgtgagcg tagctgctgt ggaaaggaac gtcctcaaaa gtgtagctga
5281 aggtaaagtt gtttccggta cgcagcatct gagaaggaaa gtactccagg cagtaaaatg
5341 aagagcgtcc tactgcctga ctcccgttgt tcagggtgag gtatccatac tgtggcacca
5401 tgaagacgtc tgctgggaac ggcgggaggc atccttgatg cgccgagccg aggacgtacg
5461 ggagctggta ctccgagtca gtaaacacct gaaccgtgct ggtaaggtta ttggcaatcg
5521 tcgtcgtacc gtcattctgc gtgacctctt tgacttgaat gttaaagagc ttgaagttga
5581 gtctcttggg tcggaatccc cagttgttgt tgatgagtct ttgccagtca cgtggtgaaa
5641 agtggcagtg gaatctgttg aagtcaaaat acccccaagg ggtgctgtag ccaaagtagt
5701 gattgtcgtt cgaggctcct gattggctgg aaatttgttt gtagaggtgg ttgttgtagg
5761 tgggcagggc ccaggttcgg gtgctggtgg tgatgactct gtcgcccatc catgtggaat
5821 cgcaatgcca atttcccgag gaattaccca ctccgtcggc gccctcgtta ttgtctgcca
5881 ttggtgcgcc actgcctgta gccatcgtat tagttcccag accagagggg gctgctggtg
5941 gctgtccgag aggctggggg tcaggtactg agtctgcgtc tccagtctga ccaaaattca
6001 atctttttct tgcaggctgc tggcccgcct ttccggttcc cgaggaggag tctggctcca
6061 caggagagtg ctctaccggc ctcttttttc ccggagccgt cttaacaggt tcctcaacca
6121 ggcccagagg ttcaagaacc ctctttttcg cctggaagac tgctcgtccg aggttgcccc
6181 caaaagacgt atcttcttta aggcgctcct gaaactccgc gtcggcgtgg ttgtacttga
6241 ggtacgggtt gtctccgctg tcgagctgcc ggtcgtaggc tttgtcgtgc tcgagggccg
6301 cggcgtctgc ctcgttgacc ggctctccct tgtcgagtcc gttgaagggt ccgaggtact
6361 tgtacccagg aagcacaaga cccctgctgt cgtccttatg ccgctctgcg ggctttggtg
6421 gtggtgggcc aggtttgagc ttccaccact gtcttattcc ttcagagaga gtgtcctcga
6481 gccaatcggg tagataaccg tcggcagccg tcttaacagg atcccccggg tgatcaagtc
6541 ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa
6601 atgatttgat aataattctt atttaactat aatatattgt gttgggttga attaaaggtc
6661 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
6721 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
6781 ggattattca taccgtccca ccatcgggcg cggatctgcc gccctggcgg ggttttacga
6841 gattgtgatt aaggtcccca gcgaccttga cgagcatctg cccggcattt ctgacagctt
6901 tgtgaactgg gtggccgaga aggagtggga gttgccgcca gattctgact tggatctgaa
6961 tctgattgag caggcacccc tgaccgtggc cgagaagctg cagcgcgact ttctgacgga
7021 gtggcgccgt gtgagtaagg ccccggaggc ccttttcttt gtgcaatttg agaagggaga
7081 gagctacttc cacttacacg tgctcgtgga aaccaccggg gtgaaatcct tagttttggg
7141 acgtttcctg agtcagattc gcgaaaaact gattcagaga atttaccgcg ggatcgagcc
7201 gactttgcca aactggttcg cggtcacaaa gaccagaaac ggcgccggag gcgggaacaa
7261 ggtggtggac gagtgctaca tccccaatta cttgctcccc aaaacccagc ctgagctcca
7321 gtgggcgtgg actaatttag aacagtattt aagcgcctgt ttgaatctca cggagcgtaa
7381 acggttggtg gcgcagcatc tgacgcacgt gtcgcagacg caggagcaga acaaagagaa
7441 tcagaatccc aattctgacg cgccggtgat cagatcaaaa acttcagcca ggtacatgga
7501 gctggtcggg tggctcgtgg acaaggggat tacctcggag aagcagtgga tccaggagga
7561 ccaggcctca tacatctcct tcaatgcggc ctccaactcg cggtcccaaa tcaaggctgc
7621 cttggacaat gcgggaaaga ttatgagcct gactaaaacc gcccccgact acctggtggg
7681 ccagcagccc gtggaggaca tttccagcaa tcggatttat aaaattttgg aactaaacgg
7741 gtacgatccc caatatgcgg cttccgtctt tctgggatgg gccacgaaaa agttcggcaa
7801 gaggaacacc atctggctgt ttgggcctgc aactaccggg aagaccaaca tcgcggaggc
7861 catagcccac actgtgccct tctacgggtg cgtaaactgg accaatgaga actttccctt
7921 caacgactgt gtcgacaaga tggtgatctg gtgggaggag gggaagatga ccgccaaggt
7981 cgtggagtcg gccaaagcca ttctcggagg aagcaaggtg cgcgtggacc agaaatgcaa
8041 gtcctcggcc cagatagacc cgactcccgt gatcgtcacc tccaacacca acatgtgcgc
8101 cgtgattgac gggaactcaa cgaccttcga acaccagcag ccgttgcaag accggatgtt
8161 caaatttgaa ctcacccgcc gtctggatca tgactttggg aaggtcacca agcaggaagt
8221 caaagacttt ttccggtggg caaaggatca cgtggttgag gtggagcatg aattctacgt
8281 caaaaagggt ggagccaaga aaagacccgc ccccagtgac gcagatataa gtgagcccaa
8341 acgggtgcgc gagtcagttg cgcagccatc gacgtcagac gcggaagctt cgatcaacta
8401 cgcagacagg taccaaaaca aatgttctcg tcacgtgggc atgaatctga tgctgtttcc
8461 ctgcagacaa tgcgagagaa tgaatcagaa ttcaaatatc tgcttcactc acggacagaa
8521 agactgttta gagtgctttc ccgtgtcaga atctcaaccc gtttctgtcg tcaaaaaggc
8581 gtatcagaaa ctgtgctaca ttcatcatat catgggaaag gtgccagacg cttgcactgc
8641 ctgcgatctg gtcaatgtgg atttggatga ctgcatcttt gaacaataaa ctcgaggaat
8701 caagcttgca tgcctgcagg tcgactctag agcctgcagt ctcgacaagc ttgtcgagaa
8761 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
8821 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
8881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
8941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
9001 tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt
9061 tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc
9121 atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt
9181 tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa
9241 acatcctgcc aactccatgt gacaaaccgt catcttcggc tacttt
//
