| 质粒名称: | pFastBac Dual His6 TEV LIC cloning vector (5B) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 30122 |
| 存储实验室: | Scott Gradia |
|
载体骨架基本信息 |
|
| 载体名称: | pFastBac Dual |
| 载体出品公司: | -- |
| 空载体大小: | 5306 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 昆虫表达 |
| 筛选标记: | 庆大霉素 |
| 载体抗性: | 氨苄青霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | His6 TEV（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | LIC site（在克隆过程中被破坏） |
| 3’克隆位点: | LIC site（在克隆过程中被破坏） |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pFastBac Dual His6 TEV LIC cloning vector (5B) was a gift from Scott Gradia (Addgene plasmid # 30122 ; http://n2t.net/addgene:30122 ; RRID:Addgene_30122) |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pFastBac Dual His6 TEV LIC cloning vector (5B) 5306 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFastBac Dual His6 TEV LIC cloning vector (5B)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5306)
TITLE 5-series pFastBac Dual LIC vectors (for insect cell expression)
REFERENCE 2 (bases 1 to 5306)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5306
/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 583..687
/gene="bla"
/label=AmpR promoter
CDS 688..1548
/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(906..925)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1719..2307
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2208..2227
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2461..2478
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 2610..2834
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(2901..3434)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDVDPSTAT"
promoter complement(3623..3651)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal 4160..4208
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4233..4252
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
promoter complement(4346..4455)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 4479..4570
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 4495..4512
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 4632..4649
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 4650..4670
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
polyA_signal 4896..5030
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4933..4952)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4987..5006
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 5059..5224
/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 ttaacaaact
541 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
661 cttcaatact attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
1021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
1081 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
1141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
1201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
1261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
1321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
1381 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
1441 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
1501 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
1561 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
1621 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
1681 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
1741 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
1801 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
1861 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
1921 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
1981 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
2041 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
2101 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
2161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
2221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
2281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
2341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
2401 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
2461 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
2521 ctgtgcggta tttcacaccg cagaccagcc gcgtaacctg gcaaaatcgg ttacggttga
2581 gtaataaatg gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac
2641 aaaatagatc taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa
2701 atcagtccag ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc
2761 ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg
2821 taaagactat attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg
2881 atctcggctt gaacgaattg ttaggtggcg gtacttgggt cgacatcaaa gtgcatcact
2941 tcttcccgta tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct
3001 tgcacgtaga tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc
3061 gcggtggcaa tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc
3121 tcactacgcg gctgctcaaa cctgggcaga acgtaagccg cgagagcgcc aacaaccgct
3181 tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa
3241 tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga
3301 tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg
3361 cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
3421 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
3481 ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa
3541 aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga
3601 gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg
3661 tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga
3721 ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc
3781 attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctgc cctggcttca
3841 ggagatcggt agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt
3901 ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg actctagcta
3961 tagttctagt ggttggccta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
4261 cccccatctc ccggtaccgc atgcttatgg agttgggata tctatatctc cttcttaaag
4321 ccgggtgatc aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt
4381 ttaattaata tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg
4441 gttgaattaa aggtccgtat actccggact attaatagat catggagata attaaaatga
4501 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
4561 acctataact attccggatt attcataccg tcccaccatc gggcgcggat cccggtccga
4621 tgggttcttc tcaccatcac catcaccatg aaaacctgta cttccaatcc aatattggaa
4681 gtggataacg gatccgaatt caaaggccta cgtcgacgag ctcactagtc gcggccgctt
4741 tcgaatctag agcctgcagt ctcgacaagc ttgtcgagaa gtactagagg atcataatca
4801 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
4861 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
4921 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
4981 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc
5041 ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt
5101 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
5161 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
5221 cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt
5281 gacaaaccgt catcttcggc tacttt
//
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