| 质粒名称: | pFUS_B3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31020 |
| 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_B3 was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31020 ; http://n2t.net/addgene:31020 ; RRID:Addgene_31020) |
| 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
pFUSB3载体基本信息
pFUSB3载体图谱质粒图谱和多克隆位点信息
pFUSB3载体简介
pFUSB3载体序列
LOCUS pFUS_B3 2991 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_B3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2991)
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 2991)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2991
/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 898..1689
/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 1782..2370
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2271..2290
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2524..2541
/label=L4440
/note="L4440 vector, forward primer"
terminator 2700..2727
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2819..2905
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2826..341)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
HLVRGQAMRRLRLAVVLQRRDWENIHASVNARVGNCQASNKTKGSVGRLGLSFYLLFVG
ERSPE"
primer_bind 2969..2985
/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 cgccaggacc atggcctgac cccggaccaa gtggtggcta tcgagacgtc
541 tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
601 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
661 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
721 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
781 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
841 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
901 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
961 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
1021 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
1081 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
1141 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
1201 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
1261 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
1321 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
1381 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
1441 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
1501 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
1561 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
1621 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
1681 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
1741 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
1801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
1861 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
1921 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
1981 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
2041 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
2101 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
2161 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
2221 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
2281 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
2341 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
2401 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
2461 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
2521 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
2581 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
2641 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
2701 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
2761 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
2821 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
2881 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
2941 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa g
//
