基本信息
| 启动子: | PgpdA, TrpC |
| 复制子: | pUC ori，F1 ori |
| 质粒分类: | 丝状真菌载体；RNAi基因沉默干扰质粒 |
| 质粒大小: | 5567bp |
| 原核抗性: | 氨苄青霉素Ampicillin |
| 筛选标记: | 新霉素Neomycin/G418 |
| 克隆菌株: | DH5α |
| 培养条件: | 37℃，有氧，LB |
| 表达宿主: | 丝状真菌 |
| 5'测序引物: | 根据序列设计 |
| 3'测序引物: | 根据序列设计 |
质粒简介
an RNA-silencing vector, pSilent-Dual1 (pSD1), with a convergent dual promoter system that provides a high-throughput platform for functional genomics research in filamentous fungi. In the pSD1 system.
质粒图谱
质粒序列
LOCUS Exported 5567 bp ds-DNA circular SYN 31-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSilient-Dual1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5567)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, August 31, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..5567
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1041..1835
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 2328..2700
/note="PtrpC"
primer_bind complement(2732..2748)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(2757..3360)
/note="PgpdA"
promoter complement(3379..3397)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3418..3434)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3442..3458
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(3466..3496)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3511..3532
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3820..4408)
/direction=LEFT
/note="pUC ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4579..5439)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5440..5544)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
661 gccccccctc gaggacgtta actgatattg aaggagcact ttttgggctt ggctggagct
721 agtggaggtc aacaatgaat gcctattttg gtttagtcgt ccaggcggtg agcacaaaat
781 ttgtgtcgtt tgacaagatg gttcatttag gcaactggtc agatcagccc cacttgtagc
841 agtagcggcg gcgctcgaag tgtgactctt attagcagac aggaacgagg acattattat
901 catctgctgc ttggtgcacg ataacttggt gcgtttgtca agcaaggtaa gtgaacgacc
961 cggtcatacc ttcttaagtt cgcccttcct ccctttattt cagattcaat ctgacttacc
1021 tattctaccc aagcatcgat atgattgaac aagatggatt gcacgcaggt tctccggccg
1081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
1141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
1201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
1261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
1321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
1381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
1441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
1501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
1561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
1621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
1681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
1741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
1801 tcgccttcta tcgccttctt gacgagttct tctgaggatc cacttaacgt tactgaaatc
1861 atcaaacagc ttgacgaatc tggatataag atcgttggtg tcgatgtcag ctccggagtt
1921 gagacaaatg gtgttcagga tctcgataag atacgttcat ttgtccaagc agcaaagagt
1981 gccttctagt gatttaatag ctccatgtca acaagaataa aacgcgtttc gggtttacct
2041 cttccagata cagctcatct gcaatgcatt aatgcattgg acctcgcaac cctagtacgc
2101 ccttcaggct ccggcgaagc agaagaatag cttagcagag tctattttca ttttcgggag
2161 acgagatcaa gcagatcaac ggtcgtcaag agacctacga gactgaggaa tccgctcttg
2221 gctccacgcg actatatatt tgtctctaat tgtactttga catgctcctc ttctttactc
2281 tgatagcttg actatgaaaa ttccgtcacc agccctgggt tctcgaggtc gacgacgtta
2341 actgagattg aaggagcact ttttgggctt ggctggagct agtggaggtc aacaatgaat
2401 gcctattttg gtttagtcgt ccaggcggtg agcacaaaat ttgtgtcgtt tgacaagatg
2461 gttcatttag gcaactggtc agatcagccc cacttgtagc agtagcggcg gcgctcgaag
2521 tgtgactctt attagcagac aggaacgagg acattattat catctgctgc ttggtgcacg
2581 ataacttggt gcgtttgtca agcaaggtaa gtgaacgacc cggtcatacc ttcttaagtt
2641 cgcccttcct ccctttattt cagattcaat ctgacttacc tattctaccc aagcatcgat
2701 aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc ggccgcaaat
2761 aaaggttctt ggatgggaag atgaatatac tgaagatggg aaaagaaaga gaaaagaaaa
2821 gagcagctgg tggggagagc aggaaaatat ggcaacaaat gttggactga cgcaacgacc
2881 ttgtcaaccc cgccgacaca ccgggcggac agacggggca aagctgccta ccagggactg
2941 agggacctca gcaggtcgag tgcagagcac cggatgggtc gactgccagc ttgtgttccc
3001 ggtctgcgcc gctggccagc tcctgagcgg cctttccggt ttcatacacc gggcaaagca
3061 ggagaggcac gatatttgga cgccctacag atgccggatg ggccaattag ggagcttacg
3121 cgccgggtac tcgctctacc tacttcggag aaggtactat ctcgtgaatc ttttaccaga
3181 tcggaagcaa ttggacttct gtacctaggt taatggcatg ctatttcgcc gacggctata
3241 cacccctggc ttcacattct ccttcgctta ctgccggtga ttcgatgaag ctccatattc
3301 tccgatgatg caatagattc ttggtcaacg aggggcacac cagcctttcc acttcggggc
3361 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
3421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
3481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
3541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
3601 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
3661 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
3721 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
3781 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
3841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
3901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
3961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
4021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
4081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
4141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
4201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
4261 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
4321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
4381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
4441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
4501 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
4561 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
4621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
4681 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
4741 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
4801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
4861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
4921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
4981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
5041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
5101 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
5161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
5221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
5281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
5341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
5401 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
5461 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
5521 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
温馨提示：不可用于临床ZL。
