基本信息
| 别名: | pFastBac Dual |
| 启动子: | P10+PH promoter |
| 复制子: | pUC ori，F1 ori |
| 终止子: | SV40 poly(A) signal |
| 质粒分类: | 广宿主系列，昆虫细胞载体 |
| 质粒大小: | 5238bp |
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | 庆大霉素Gen |
| 克隆菌株: | 大肠杆菌DH5α |
| 培养条件: | 37℃，有氧 LB |
| 表达宿主: | 昆虫细胞 |
| 诱导方式: | 无须诱导，瞬时表达 |
| 5'测序引物: | pFastbac-F: TATTCCGGATTATTCATACC |
| 3'测序引物: | pFastBac-R: ACAAATGTGGTATGGCTGA |
质粒简介
pFastBac Dual昆虫细胞表达载体包含有两个启动子，能够同时表达两个外源基因，一个表达基因是polyhedrin (PH)启动子控制，另一个外源基因是由p10启动子控制。
pFastBacDual 载体的特点是在单个载体上有两个启动子，用于在使用 Bac-to-Bac® 杆状病毒表达系统(Bac-to-Bac® Baculovirus Expression System) 时在昆虫细胞中同时表达两个蛋白质。该载体具有两个强启动子，多角体蛋白启动子和p10 启动子，因此可实现高水平表达。包含 CAT和β-葡萄糖醛酸酶 (gus) 的表达对照。
The Bac-to-Bac® Vector Kit contains a pFastBac™ 1 vector, as well an expression control vector, intended for use as part of the Bac-to-Bac® Baculovirus Expression System , which enables the efficent production of recombinant baculovirus for expression testing in insect cells. The Bac-to-Bac® System relies on generation of recombinant baculovirus by site-specific transposition in E. colirather than homologous recombination in insect cells (Figure 1). This system features:
Time-saving expression bacmid. With Bac-to-Bac®, the expression cassette of the pFastBac™ vector recombines with the parent bacmid in DH10Bac™ E. coli Competent Cells (not included with this vector kit) to form an expression bacmid. The bacmid is then transfected into insect cells for production of recombinant baculovirus particles.
Easy colony screening. The parent bacmid in DH10Bac™ E. colicontains a segment of the lacZα gene. The lacZα gene is disrupted upon transposition of the expression cassette into the bacmid allowing for blue/white selection of recombinants. This makes identification of recombinant colonies easy.质粒图谱
质粒序列
LOCUS Exported 5238 bp ds-DNA circular SYN 13-1-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5238)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016-1-13
FEATURES Location/Qualifiers
source 1..5238
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 102..557
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 584..688
/gene="bla"
/note="AmpR promoter"
CDS 689..1549
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1720..2308
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
mobile_element 2611..2835
/mobile_element_type="transposon:Tn7"
/note="Tn7R"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2902..3435)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/note="GmR"
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(3624..3652)
/gene="intI1 (promoter lies within the coding sequence)"
/note="Pc promoter"
/note="class 1 integron promoter"
polyA_signal 4161..4209
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
misc_feature 4274..4337
/note="MCS1"
promoter complement(4345..4454)
/note="p10 promoter"
/note="baculovirus promoter for expression in insect cells"
promoter 4478..4569
/gene="polh from Autographa californica"
/note="polyhedrin promoter"
/note="promoter for the baculovirus polyhedrin gene"
misc_feature 4606..4704
/note="MCS2"
polyA_signal 4828..4962
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
mobile_element 4991..5156
/mobile_element_type="transposon:Tn7"
/note="Tn7L"
/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 agcaccatgg ctcgagatcc
4321 cgggtgatca agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt
4381 taattaatat acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg
4441 ttgaattaaa ggtccgtata ctccggaata ttaatagatc atggagataa ttaaaatgat
4501 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
4561 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc ccggtccgaa
4621 gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc tttcgaatct
4681 agagcctgca gtctcgacaa gcttgtcgag aagtactaga ggatcataat cagccatacc
4741 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
4801 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
4861 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
4921 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gcttgagcct
4981 aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
5041 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
5101 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
5161 ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat gtgacaaacc
5221 gtcatcttcg gctacttt
//
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