| 质粒名称: | pFUS_B10 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31027 |
| 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_B10 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31027 ; http://n2t.net/addgene:31027 ; RRID:Addgene_31027) |
| 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
pFUSB10载体基本信息
pFUSB10载体图谱质粒图谱和多克隆位点信息
pFUSB10载体简介
pFUSB10载体序列
LOCUS pFUS_B10 3041 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B10
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3041)
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 3041)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3041
/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 1037..1828
/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 1921..2509
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2410..2429
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2663..2680
/label=L4440
/note="L4440 vector, forward primer"
terminator 2839..2866
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2958..3
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2965..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 tggcggcaag caagcgctcg
601 aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag
661 tggtggctat cgagacgtct agaccagcca ggacagaaat gcctcgactt cgctgctacc
721 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
781 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
841 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
901 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
961 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
1021 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
1081 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
1141 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
1201 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
1261 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
1321 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
1381 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
1441 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
1501 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
1561 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
1621 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
1681 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
1741 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
1801 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
1861 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
2041 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
2281 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
2521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
2581 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
2641 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
2701 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
2761 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
2821 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
2881 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
2941 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
3001 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta t
//
