| 质粒名称: | pFUS_B2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31019 |
| 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_B2 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31019 ; http://n2t.net/addgene:31019 ; RRID:Addgene_31019) |
| 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
pFUSB2载体基本信息
pFUSB2载体图谱质粒图谱和多克隆位点信息
pFUSB2载体简介
pFUSB2载体序列
LOCUS pFUS_B2 2988 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2988)
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 2988)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2988
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 206..222
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 232..251
/label=T7
/note="T7 promoter, forward primer"
promoter 232..250
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(298..318)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(298..316)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(337..353)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(337..353)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(350..372)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 361..377
/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(385..415)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 430..451
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 895..1686
/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 1779..2367
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2268..2287
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2521..2538
/label=L4440
/note="L4440 vector, forward primer"
terminator 2697..2724
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2816..2902
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2823..341)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG
ERSPE"
primer_bind 2966..2982
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 cgtctcatgg cctgaccccg gaccaagtgg tggctatcga gaccggcgcc gctacagggc
61 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
121 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
181 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
241 tcactatagg gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct
301 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
481 ccggtggtct caggaccatg gcctgacccc ggaccaagtg gtggctatcg agacgtctag
541 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
601 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
661 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
721 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
781 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
841 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
901 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
961 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
1021 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
1081 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
1141 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
1201 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
1261 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
1321 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
1381 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
1441 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
1501 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
1561 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
1621 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
1681 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
1741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
1801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
1861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
1921 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
1981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
2041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
2101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
2161 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
2221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
2281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
2341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
2401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
2461 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
2521 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
2581 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
2641 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
2701 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
2761 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
2821 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
2881 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
2941 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaag
//
