| 质粒名称: | pFUS_B5 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31022 |
| 存储实验室: | Adam Bogdanove, Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pCR8 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2447 bp |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | LacZ + BsaI restriction sites |
| 插入大小: | 459 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AflII (未破坏) |
| 3' 克隆位点: | XbaI(未破坏) |
| 5' 测序引物 | pCR8_F1 (ttgatgcctggcagttccct) |
| 3' 测序引物 | pCR8_R1 (cgaaccgaacaggcttatgt) |
|
文献引用方法 |
|
| 材料和方法部分: | pFUS_B5 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31022 ; http://n2t.net/addgene:31022 ; RRID:Addgene_31022) |
| 参考文献部分: | Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. ():. 10.1093/nar/gkr218 PubMed 21493687 |
买家导航
pFUSB5载体基本信息
pFUSB5载体图谱质粒图谱和多克隆位点信息
pFUSB5载体简介
pFUSB5载体序列
LOCUS pFUS_B5 3016 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3016)
AUTHORS Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller
JA, Somia NV, Bogdanove AJ, Voytas DF
TITLE Efficient design and assembly of custom TALEN and other TAL
effector-based constructs for DNA targeting.
JOURNAL Nucleic Acids Res. 2011 Apr 14. ():.
PUBMED 21493687
REFERENCE 2 (bases 1 to 3016)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3016
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 67..83
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 295..311
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 321..340
/label=T7
/note="T7 promoter, forward primer"
promoter 321..339
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(387..407)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(387..405)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(426..442)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(426..442)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(439..461)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 450..466
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(474..504)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 519..540
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 1012..1803
/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 1896..2484
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2385..2404
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2638..2655
/label=L4440
/note="L4440 vector, forward primer"
terminator 2814..2841
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2933..3
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2940..430)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG
ERSPE"
ORIGIN
1 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
61 gacgttgtaa aacgacggcc agtcttaagc gtctcatggc ctgaccccgg accaagtggt
121 ggctatcgag accggcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
181 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
241 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
301 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
361 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
541 attgcgttgc gctcactgcc cgctttccac cggtggtctc gtgcagcggc tgttgccggt
601 gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgaga cgtctagacc
661 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
721 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
781 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
841 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
901 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
961 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
1021 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
1081 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
1141 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
1201 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
1261 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
1321 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
1381 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
1441 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
1501 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
1561 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
1621 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
1681 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
1741 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
1801 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
1861 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
1921 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
1981 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
2041 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
2101 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
2161 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
2221 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
2281 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
2341 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
2401 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
2461 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
2521 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
2581 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
2641 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
2701 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
2761 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
2821 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
2881 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
2941 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
3001 gagcctttcg ttttat
//
