| 质粒名称: | pFUS_B4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31021 |
| 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_B4 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31021 ; http://n2t.net/addgene:31021 ; RRID:Addgene_31021) |
| 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
pFUSB4载体基本信息
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LOCUS pFUS_B4 3007 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3007)
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 3007)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3007
/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 1003..1794
/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 1887..2475
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2376..2395
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2629..2646
/label=L4440
/note="L4440 vector, forward primer"
terminator 2805..2832
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2924..3
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2931..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 ctgttgccgg tgctgtgcca
601 ggaccatggc ctgaccccgg accaagtggt ggctatcgag acgtctagac cagccaggac
661 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
721 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
781 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
841 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
901 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
961 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
1021 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
1081 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
1141 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
1201 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
1261 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
1321 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
1381 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
1441 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
1501 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
1561 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
1621 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
1681 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
1741 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
1801 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
1861 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
1921 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
1981 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
2041 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
2101 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
2161 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
2221 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
2281 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
2341 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
2401 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
2461 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
2521 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
2581 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
2641 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
2701 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
2761 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
2821 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
2881 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
2941 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
3001 gttttat
//
