| 质粒名称: | pBMCL1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178203 |
| 存储实验室: | Lei Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pFastBac1 |
| 载体出品公司: | Thermofisher |
| 空载体大小: | 4775 bp |
| 载体修饰: | 为哺乳动物细胞中的蛋白质表达添加多种元素 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | ggtcttactgacatc |
| 3’测序引物 | ccagtcaatctttcac |
|
文献引用方法 |
|
| 材料和方法部分: | pBMCL1 was a gift from Lei Chen (Addgene plasmid # 178203 ; http://n2t.net/addgene:178203 ; RRID:Addgene_178203) |
| 参考文献部分: | An improved vector for baculovirus-mediated protein production in mammalian cells. Wenjun G, Wang M, Chen L. bioRxiv 2021.10.18.464913 10.1101/2021.10.18.464913 |
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 pBMCL1 7124 bp ds-DNA circular SYN 06-DEC-2021
DEFINITION An improved vector for BacMam system.
ACCESSION .
VERSION .
KEYWORDS pBMCL1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7124)
AUTHORS Wenjun G, Wang M, Chen L
TITLE An improved vector for baculovirus-mediated protein production in
mammalian cells
JOURNAL bioRxiv 2021.10.18.464913
REFERENCE 2 (bases 1 to 7124)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 6, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7124
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 581..605
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 609..741
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 724..743
/label=pMT2-F
/note="Synthetic intron, forward primer"
CDS 782..799
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 806..1394
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(859..879)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1277..1288)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 1521..1655
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1558..1577)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1612..1631
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 2054..2082
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
mobile_element 2852..3076
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
primer_bind complement(3210..3227)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3381..3969)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3461..3480)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4140..5000)
/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 4763..4782
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5001..5105)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5131..5585)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5268..5289)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5479..5498
/label=F1ori-R
/note="F1 origin, reverse primer"
mobile_element 5769..5934
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
polyA_signal 6132..6180
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 6205..6224
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(6635..6667)
/codon_start=1
/product="epitope tag from vesicular stomatitis virus G
protein"
/label=VSV-G tag
/translation="YTDIEMNRLGK"
promoter complement(7001..7092)
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind complement(7059..7076)
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
ORIGIN
1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agatcaaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
661 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
721 actttgcctt tctctccaca ggtgtaagga attcggatcc cgggcggccg ctagcctcga
781 gcaccaccac caccaccact gagataatca acctctggat tacaaaattt gtgaaagatt
841 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
901 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
961 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
1021 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
1081 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
1141 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
1201 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
1261 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
1321 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
1381 ggccgcctcc ccgcctggtc gagaagtact agaggatcat aatcagccat accacatttg
1441 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
1501 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
1561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
1621 ccaaactcat caatgtatct tatcatgtct ggatctgatc agtaacaaca ccatttaaat
1681 ggagtggtta caaatggagt ggttaattaa gttccggagt atacgtagcc aaccactaga
1741 actatagcta gagtcctggg cgaacaaacg atgctcgcct tccagaaaac cgaggatgcg
1801 aaccacttca tccggggtca gcaccaccgg caagcgccgc gacggccgag gtcttccgat
1861 ctcctgaagc cagggcagat ccgtgcacag caccttgccg tagaagaaca gcaaggccgc
1921 caatgcctga cgatgcgtgg agaccgaaac cttgcgctcg ttcgccagcc aggacagaaa
1981 tgcctcgact tcgctgctgc ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
2041 ggcacgaacc cagttgacat aagcctgttc ggttcgtaaa ctgtaatgca agtagcgtat
2101 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
2161 ggttttcatg gcttgttatg actgtttttt tgtacagtct atgcctcggg catccaagca
2221 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
2281 gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg
2341 taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga
2401 gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt
2461 gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc
2521 tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga
2581 tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct
2641 caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga
2701 cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga
2761 tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc
2821 ggagttgttc ggtaaattgt cacaacgccg cgaatatagt ctttaccatg cccttggcca
2881 cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt tgctttagcc
2941 ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag tttacaacta
3001 ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca gtttaagact
3061 ttattgtccg cccacacccg cttacgcagg gcatccattt attactcaac cgtaaccgat
3121 tttgccaggt tacgcggctg gtctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
3181 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
3241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
3301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
3361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
3421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
3481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
3541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
3601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
3661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
3721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
3781 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
3841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
3901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
3961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
4081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
4141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
4201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
4261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
4321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
4381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
4441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
4501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
4561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
4621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
4681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
4741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
4801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
4861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
4921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
4981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
5041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
5101 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
5161 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
5221 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
5281 agagtccact attaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
5341 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
5401 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
5461 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
5521 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
5581 cgcgtcccat tcgccattca ggctgcaaat aagcgttgat attcagtcaa ttacaaacat
5641 taataacgaa gagatgacag aaaaattttc attctgtgac agagaaaaag tagccgaaga
5701 tgacggtttg tcacatggag ttggcaggat gtttgattaa aaacataaca ggaagaaaaa
5761 tgccccgctg tgggcggaca aaatagttgg gaactgggag gggtggaaat ggagttttta
5821 aggattattt agggaagagt gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa
5881 tacgaaaatt aaaaatgaca aaatagtttg gaactagatt tcacttatct ggttcggatc
5941 tcctaggctc aagcagtgat cagtggctat ggcagggctt gccgccccga cgttggctgc
6001 gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag aaacgcgggc
6061 gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
6121 cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt tttattgccg
6181 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctcccccatc
6241 tcccgctcta gtacttctcg acaagctttt aggccagggc tttaccctcg gtcttggagg
6301 tgacgtccac ctggacaccc acgaacttgg acacgcgacc gtcgggagtc ttgatggggg
6361 tcacagtcag caggttccag aaaggggtac cgtccttgcg gtagttcagc agacgcacgg
6421 agcaggcttc acccttcttg atagcgtcgc ggatcttctg cacctccttg gggtcggtac
6481 cttcaccctg caggaagcgg gcgttgtggc ccagcacttc gtcggggccg taaccagtca
6541 tggcgtagaa gccctcggaa gcgtacacca gggggcagtc gggcagagta gcgtcagcca
6601 ccacgaaggt gtgacgcaga ccggcaccgc cacctttacc caggcggttc atctcgatgt
6661 cagtatatat ctgacgcttc ttggtgtgct tcagcttgat gcacaggtgg atacccacgc
6721 gcaggaccag gaacagaccg atgatcaggc cgatgatgaa gaagaaagaa gcgatgctgg
6781 acttccagct gctgaaccag ccttcgacca gttcgatagg gttcttgctc agaccggtgt
6841 caccagcgaa ggcggagtga gcagcagcag ccagcagcac gtacagcacg atagcgctca
6901 ccatcttgct agtgtgctcc ttgttgaaac cctggtggct ctggttcacc agcagcatag
6961 atccgcgccc gatggtggga cggtatgaat aatccggaat atttataggt ttttttatta
7021 caaaactgtt acgaaaacag taaaatactt atttatttgc gagatggtta tcattttaat
7081 tatctccatg atctattaat acttaattaa caacaccatt tgtt
//
