| 质粒名称: | pBIG1a |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80611 |
| 存储实验室: | Jan-Michael Peters |
|
载体骨架基本信息 |
|
| 载体名称: | pBIG1a |
| 载体出品公司: | -- |
| 空载体大小: | 5915 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: | 氨苄霉素和大观霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
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: | -- |
| 克隆方法: | Gibson克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | TCAACAGGTTGAACTGCTGATC |
| 3’测序引物 | GGTGTAGCGTCGTAAGCTAATAC |
|
文献引用方法 |
|
| 材料和方法部分: | pBIG1a was a gift from Jan-Michael Peters (Addgene plasmid # 80611 ; http://n2t.net/addgene:80611 ; RRID:Addgene_80611) |
| 参考文献部分: | biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes. Weissmann F, Petzold G, VanderLinden R, Huis In 't Veld PJ, Brown NG, Lampert F, Westermann S, Stark H, Schulman BA, Peters JM. Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2564-9. doi: 10.1073/pnas.1604935113. Epub 2016 Apr 25. 10.1073/pnas.1604935113 PubMed 27114506 |
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 pBIG1a 5915 bp ds-DNA circular SYN 12-JUL-2021
DEFINITION pBIG1a - Step1 vector a (biGBac system for expression of protein
complexes in insect cells).
ACCESSION .
VERSION .
KEYWORDS pBIG1a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5915)
AUTHORS Weissmann F, Petzold G, VanderLinden R, Huis In 't Veld PJ, Brown
NG, Lampert F, Westermann S, Stark H, Schulman BA, Peters JM
TITLE biGBac enables rapid gene assembly for the expression of large
multisubunit protein complexes.
JOURNAL Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2564-9. doi:
10.1073/pnas.1604935113. Epub 2016 Apr 25.
PUBMED 27114506
REFERENCE 2 (bases 1 to 5915)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5915
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 442..1233
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 1274..1728
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1361..1380)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1570..1591
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1754..1858
/gene="bla"
/label=AmpR promoter
CDS 1859..2719
/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(2077..2096)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2890..3478
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3379..3398
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3632..3649
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 3783..4007
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS complement(4074..4607)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(4796..4824)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 5220..5253
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(5299..5318)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5343..5391
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
mobile_element 5668..5833
/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 ctgggaggca gaataaatga tcatatcgtc
121 aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg agaagcacac
181 ggtcacactg cttccggtag tcaataaacc ggtaagtagc gtatgcgctc acgcaactgg
241 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
301 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
361 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
421 cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta
481 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
541 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
601 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
661 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
721 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
781 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
841 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
901 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
961 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
1021 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
1081 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
1141 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
1201 aaaggcgaga tcaccaaggt agtcggcaaa taatgtctaa caattcgttc aagccgacgg
1261 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
1321 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
1381 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
1441 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
1501 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
1561 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
1621 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
1681 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
1741 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
1801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
1861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
1921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
1981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
2041 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
2101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
2161 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
2221 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
2281 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
2341 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
2401 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
2461 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
2521 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
2581 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
2641 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
2701 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
2761 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
2821 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
2881 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
2941 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
3001 aactggcttc agcagagcgc agataccaaa tactattctt ctagtgtagc cgtagttagg
3061 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
3121 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
3181 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
3241 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
3301 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
3361 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
3421 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
3481 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
3541 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
3601 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
3661 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca
3721 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg
3781 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa
3841 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc
3901 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg
3961 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac
4021 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
4081 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
4141 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
4201 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
4261 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
4321 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
4381 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
4441 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
4501 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
4561 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
4621 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
4681 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
4741 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
4801 acgaaccgaa caggcttatg tcaactgggt ttgtgccttc atccgtttcc acggtgtgcg
4861 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
4921 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
4981 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg
5041 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
5101 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacagctttg
5161 tttgtactat caacaggttg aactgctgat caacagatcc tctacgcggc cgcggtacca
5221 taacttcgta tagcatacat tatacgaagt tatctgggtt taaacgctct atggtctaaa
5281 gtttttatcg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
5341 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
5401 gggttcggtc ccagggctgg cactctgtca agtgaacgct ctatggtcta aagatttaaa
5461 tctatatctc aatcggggtt acgtcgatac cccaccgaga ccccattggg accaatacgc
5521 ccgcgtttct tccttttccc caccccaacc cccaagttcg ggtgaaggcc cagggctcgc
5581 agccaacgtc ggggcggcaa gccctgccat agccactacg ggttaattaa aaactatatc
5641 tcaatcgggg tttaaacagg agatccgaac cagataagtg aaatctagtt ccaaactatt
5701 ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca tctattttgt
5761 cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt
5821 ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca
5881 actccatgtg acaaaccgtc atcttcggct acttt
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：Tol2-elavl3-H2B-GCaMP6s
- 下一篇：pX037
