| 质粒名称: | pFUS_A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31028 |
| 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_A was a gift from Adam Bogdanove & Daniel Voytas (Addgene plasmid # 31028 ; http://n2t.net/addgene:31028 ; RRID:Addgene_31028) |
| 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
pFUSA载体基本信息
pFUSA载体图谱质粒图谱和多克隆位点信息
pFUSA载体简介
pFUSA载体序列
LOCUS pFUS_A 3019 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFUS_A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3019)
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 3019)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3019
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(57..434)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH
HLVRGQVQGETLKTGRRFTTS"
primer_bind 67..83
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 299..315
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 325..344
/label=T7
/note="T7 promoter, forward primer"
promoter 325..343
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(391..411)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(391..409)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(430..446)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(430..446)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(443..465)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 454..470
/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(478..508)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 523..544
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 1015..1806
/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 1899..2487
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2388..2407
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2641..2658
/label=L4440
/note="L4440 vector, forward primer"
terminator 2817..2844
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2936..3
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
61 gacgttgtaa aacgacggcc agtcttaagc gtctccccct gaacctgacc ccggaccaag
121 tggtggctat cgagaccggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
181 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
241 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
301 aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg
361 ccccccctcg aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
541 attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg tctctggcgg caagcaagcg
601 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggccg agacgtctag
661 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
721 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
781 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
841 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
901 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
961 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
1021 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
1081 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
1141 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
1201 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
1261 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
1321 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
1381 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
1441 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
1501 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
1561 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
1621 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
1681 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
1741 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
1801 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
1861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
1921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
1981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
2041 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
2101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
2161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
2221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
2281 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
2341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
2401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
2461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
2521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
2581 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
2641 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
2701 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
2761 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
2821 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
2881 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
2941 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
3001 actgagcctt tcgttttat
//
