| 质粒名称: | pFGC5941-PacI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 44020 |
| 存储实验室: | Guiliang Tang |
|
载体骨架基本信息 |
|
| 载体名称: | pFGC5941 |
| 空载体大小: | 11405 bp |
| 完整质粒大小: | 8610bp |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
文献引用方法 |
|
| 材料和方法部分: | pFGC5941-PacI was a gift from Guiliang Tang (Addgene plasmid # 44020 ; http://n2t.net/addgene:44020 ; RRID:Addgene_44020) |
| 参考文献部分: | Construction of short tandem target mimic (STTM) to block the functions of plant and animal microRNAs. Tang G, Yan J, Gu Y, Qiao M, Fan R, Mao Y, Tang X. Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006. Epub 2012 Oct 23. 10.1016/j.ymeth.2012.10.006 PubMed 23098881 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pFGC5941-PacI 8610 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pFGC5941-PacI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8610)
AUTHORS Tang G, Yan J, Gu Y, Qiao M, Fan R, Mao Y, Tang X
TITLE Construction of short tandem target mimic (STTM) to block the
functions of plant and animal microRNAs.
JOURNAL Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006.
Epub 2012 Oct 23.
PUBMED 23098881
REFERENCE 2 (bases 1 to 8610)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8610
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..25
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator 111..363
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(373..924)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(930..1310)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
terminator 1380..2087
/label=OCS terminator
/note="octopine synthase terminator"
primer_bind complement(2112..2129)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2112..2128)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2121..2143)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 2331..2355
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 3656..4285
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 4714..5787
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 5853..6047
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(6283..6305)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 6391..6531
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 6446..6465
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(6546..6563)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6717..7305)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6797..6816)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7392..8186)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
61 gacgttttta atgtactgaa ttaacgccga attaattcga gctcggatct gataatttat
121 ttgaaaattc ataagaaaag caaacgttac atgaattgat gaaacaatac aaagacagat
181 aaagccacgc acatttagga tattggccga gattactgaa tattgagtaa gatcacggaa
241 tttctgacag gagcatgtct tcaattcagc ccaaatggca gttgaaatac tcaaaccgcc
301 ccatatgcag gagcggatca ttcattgttt gtttggttgc ctttgccaac atgggagtcc
361 aagattctgc agtcaaatct cggtgacggg caggaccgga cggggcggta ccggcaggct
421 gaagtccagc tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg
481 cagcatgccg cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg
541 cagcccgatg acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg
601 ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg ggggagacgt acacggtcga
661 ctcggccgtc cagtcgtagg cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc
721 gacctcgccg tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc
781 gtccgtccac tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat
841 gtagtggttg acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc
901 ggccgggcgt cgttctgggc tcatcgattc gatttggtgt atcgagattg gttatgaaat
961 tcagatgcta gtgtaatgta ttggtaattt gggaagatat aataggaagc aaggctattt
1021 atccatttct gaaaaggcga aatggcgtca ccgcgagcgt cacgcgcatt ccgttcttgc
1081 tgtaaagcgt tgtttggtac acttttgact agcgaggctt ggcgtgtcag cgtatctatt
1141 caaaagtcgt taatggctgc ggatcaagaa aaagttggaa tagaaacaga atacccgcga
1201 aattcaggcc cggttgccat gtcctacacg ccgaaataaa cgaccaaatt agtagaaaaa
1261 taaaaactga ctcggatact tacgtcacgt cttgcgcact gatttgaaaa atctcagaat
1321 tcatggatcc taggtgagtc tagagagtta attaagaccc gggactagtc cctagagtcc
1381 tgctttaatg agatatgcga gacgcctatg atcgcatgat atttgctttc aattctgttg
1441 tgcacgttgt aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca
1501 ttctaatgaa tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat
1561 ttactgattg taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc
1621 tgaataggtt tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta
1681 ttattacaaa tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta
1741 cataaatctt attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc
1801 ggcgaactaa taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag
1861 attccttgaa gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc
1921 cggtccagca cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt
1981 tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt
2041 gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac ctgcaggcat
2101 gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
2161 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
2221 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcta gagcagcttg
2281 agcttggatc agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt tgacaggata
2341 tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg
2401 gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct
2461 cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt
2521 tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct
2581 gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat
2641 acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg
2701 ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac
2761 gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg
2821 gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta
2881 gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc
2941 ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc
3001 atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc
3061 acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc
3121 ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg
3181 aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc
3241 agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg
3301 accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac
3361 cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga
3421 tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg ctgcatgaaa
3481 tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag
3541 aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg
3601 cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa
3661 ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct
3721 agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg
3781 cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga
3841 ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga
3901 cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct
3961 gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt
4021 taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc
4081 gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc
4141 cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac
4201 aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc
4261 tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca
4321 aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc
4381 ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc
4441 aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac
4501 atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc
4561 taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat
4621 gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa
4681 tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg
4741 gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc
4801 gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct
4861 gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag
4921 ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc
4981 cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga
5041 gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg
5101 gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc
5161 gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca
5221 cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac
5281 ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag
5341 gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag
5401 atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg
5461 taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt
5521 ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag
5581 gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc
5641 aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat
5701 ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc
5761 gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta
5821 gcaggggaaa aaggtcgaaa aggtctcttt cctgtggata gcacgtacat tgggaaccca
5881 aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac
5941 cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac
6001 tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag
6061 cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc
6121 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
6181 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
6241 caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
6301 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
6361 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
6421 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
6481 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
7141 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
7261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
7321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
7381 cattctaggt actaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca
7441 ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc
7501 ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca
7561 agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg
7621 ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg
7681 cgcggatctt taaatggagt gtcttcttcc cagttttcgc aatccacatc ggccagatcg
7741 ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa
7801 tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt
7861 tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg
7921 agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt
7981 ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac
8041 cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta
8101 gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat
8161 tccggtgata ttctcatttt agccatttat tatttccttc ctcttttcta cagtatttaa
8221 agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc cttgcattct
8281 aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg gcgtataaca
8341 tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg ctctgtcatc
8401 gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt
8461 tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct
8521 cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
8581 ctgccggtcg gggagctgtt ggctggctgg
//
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