| 质粒名称: | pFUS_B8 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31025 |
| 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_B8 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31025 ; http://n2t.net/addgene:31025 ; RRID:Addgene_31025) |
| 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
pFUSB8载体基本信息
pFUSB8载体图谱质粒图谱和多克隆位点信息
pFUSB8载体简介
pFUSB8载体序列
LOCUS pFUS_B8 3025 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B8
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3025)
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 3025)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3025
/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 1021..1812
/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 1905..2493
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2394..2413
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2647..2664
/label=L4440
/note="L4440 vector, forward primer"
terminator 2823..2850
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2942..3
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2949..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 ctcgaaacgg tgcagcggct
601 gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgagac
661 gtctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga
721 cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt
781 aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
841 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta
901 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
961 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt
1021 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
1081 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
1141 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
1201 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
1261 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
1321 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
1381 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
1441 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
1501 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
1561 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
1621 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
1681 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
1741 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
1801 gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg
1861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
1921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
1981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
2041 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
2101 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
2161 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
2221 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
2281 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
2341 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
2401 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
2461 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
2521 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
2581 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
2641 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
2701 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
2761 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
2821 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
2881 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
2941 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
3001 cttccgactg agcctttcgt tttat
//
