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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Plasmid#86198
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pYPQ202植物表达质粒
别称: Plasmid#86198
质粒属性
质粒简介
pYPQ202是一个植物细胞Gateway系列表达质粒。
质粒图谱
质粒序列
LOCUS Exported 12807 bp ds-DNA circular SYN 12-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12807
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1125..1319
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 1663..1803
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(1989..2577)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2664..3458)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 5082..5106
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 5184..5358
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(5399..6424)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(6491..7167)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
terminator 7489..7741
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 7805..7929
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(7970..8275)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLS-DK
VPRELYPVVHIGDESWRMMTTD-MASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(8595..9275)
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
promoter complement(9329..9359)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind complement(9384..9508)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature complement(9547..10181)
/label=AtUBQ10 promoter
misc_feature 10410..10434
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 11735..12364
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
1 ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag
61 gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg
121 caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg
181 tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat
241 gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag
301 cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt
361 tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt
421 tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc
481 gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag
541 cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag
601 cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt
661 agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta
721 gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac
781 cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc
841 gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc
901 gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg
961 ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac
1021 cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg
1081 caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac
1141 attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg
1201 tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt
1261 tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa
1321 ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc
1381 tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct
1441 ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc
1501 cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
1561 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
1621 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
1681 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
1741 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
1801 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
1861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
1921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
1981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
2041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
2101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
2161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
2221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
2281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
2341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
2401 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
2461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
2521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
2581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
2641 attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt
2701 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
2761 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
2821 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
2881 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
2941 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
3001 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
3061 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
3121 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
3181 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
3241 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
3301 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
3361 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
3421 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
3481 tacagtattt aaagataccc caagaagcta aggaaggtgc gaataagcgg ggaaattctt
3541 ctcggctgac tcagtcattt catttcttca tgtttgagcc gattttttct cccgtaaatg
3601 ccttgaatca gcctatttag accgtttctt cgccatttaa ggcgttatcc ccagttttta
3661 gtgagatctc tcccactgac gtatcatttg gtccgcccga aacaggttgg ccagcgtgaa
3721 taacatcgcc agttggttat cgtttttcag caaccccttg tatctggctt tcacgaagcc
3781 gaactgtcgc ttgatgatgc gaaatgggtg ctccaccctg gcccggatgc tggctttcat
3841 gtattcgatg ttgatggccg ttttgttctt gcgtggatgc tgtttcaagg ttcttacctt
3901 gccggggcgc tcggcgatca gccagtccac atccacctcg gccagctcct cgcgctgtgg
3961 cgccccttgg tagccggcat cggctgagac aaattgctcc tctccatgca gcagattacc
4021 cagctgattg aggtcatgct cgttggccgc ggtggtgacc aggctgtggg tcaggccact
4081 cttggcatcg acaccaatgt gggccttcat gccaaagtgc cactgattgc ctttcttggt
4141 ctgatgcatc tccggatcgc gttgctgctc tttgttcttg gtcgagctgg gtgcctcaat
4201 gatggtggca tcgaccaagg tgccttgagt catcatgacg cctgcttcgg ccagccagcg
4261 attgatggtc ttgaacaatt ggcgggccag ttgatgctgc tccagcaggt ggcggaaatt
4321 catgatggtg gtgcggtccg gcaaggcgct atccagggat aaccgggcaa acagacgcat
4381 ggaggcgatt tcgtacagag catcttccat cgcgccatcg ctcaggttgt accaatgctg
4441 catgcagtga atgcgtagca tggtttccag cggataaggt cgccggccat taccagcctt
4501 ggggtaaaac ggctcgatga cttccaccat gttttgccat ggcagaatct gctccatgcg
4561 ggacaagaaa atctcttttc tggtctgacg gcgcttactg ctgaattcac tgtcggcgaa
4621 ggtaagttga tgactcatga tgaaccctgt tctatggctc cagatgacaa acatgatctc
4681 atatcaggga cttgttcgca ccttccctaa ttataacaag acgaactcca attcactgtt
4741 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
4801 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
4861 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
4921 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
4981 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
5041 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
5101 gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactga
5161 attaacgccg aattaattcg ggggatctgg attttagtac tggattttgg ttttaggaat
5221 tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg gtttcttata
5281 tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg gaactactca
5341 cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg catctactct
5401 atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta
5461 cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc
5521 cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat
5581 tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga
5641 gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca
5701 tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga
5761 acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt
5821 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
5881 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
5941 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
6001 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
6061 tcgcatccat agcctccgcg accggttgta gaacagcggg cagttcggtt tcaggcaggt
6121 cttgcaacgt gacaccctgt gaacggcggg agatgcaata ggtcaggctc tcgctaaact
6181 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
6241 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
6301 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga
6361 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
6421 tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt gtagagagag
6481 actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt
6541 cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat
6601 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg
6661 ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat
6721 cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac
6781 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct
6841 caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt
6901 cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
6961 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
7021 tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt
7081 tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
7141 gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg
7201 atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg ctctagccaa
7261 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
7321 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
7381 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
7441 gataacaatt tcacacagga aacagctatg acatgattac gaattcccga tctagtaaca
7501 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
7561 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
7621 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
7681 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
7741 cggggaaatt cgagctccac cgcggtggcg gccgctctag aactagttaa ttaagaatta
7801 tcgaaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa atatcaatat
7861 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
7921 gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
7981 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
8041 cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
8101 catccccgat atgcaccacc gggtaaagtt cacgggggac tttatctgac agcagacgtg
8161 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
8221 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
8281 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
8341 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
8401 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
8461 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
8521 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
8581 ctgttatctg gcttttagta agccggatcc tctagattac gccccgcctg ccactcatcg
8641 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
8701 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
8761 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
8821 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
8881 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
8941 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
9001 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
9061 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
9121 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
9181 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
9241 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
9301 ctcgacggat cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag
9361 cctggggtgc ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg
9421 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
9481 ctcgttcagc ttttttgtac aaacttgttt gatagcttgg cgcgcctcga gggggggccc
9541 ggtaccctgt taatcagaaa aactcagatt aatctacaaa ttcgatcgca caaactagaa
9601 actaacacca gatctagata gaaatcacaa atcgaagagt aattattcga caaaactcaa
9661 attatttgaa caaatcggat gatatctatg aaaccctaat cgagaattaa gatgatatct
9721 aacgatcaaa cccagaaaat cgtcttcgat ctaagattaa cagaatctaa accaaagaac
9781 atatacgaaa ttgggatcga acgaaaacaa aatcgaagat tttgagagaa taaggaacac
9841 agaaatttac cttgatcacg gtagagagaa ttgagagaaa gtttttaaga ttttgagaaa
9901 ttgaaatctg aattgtgaag aagaagagct ctttgggtat tgttttatag aagaagaaga
9961 agaaaagacg aggacgacta ggtcacgaga aagctaaggc ggtgaagcaa tagctaataa
10021 taaaatgaca cgtgtattga gcgttgttta cacgcaaagt tgtttttggc taattgcctt
10081 atttttaggt tgaggaaaag tatttgtgct ttgagttgat aaacacgact cgtgtgtgcc
10141 ggctgcaacc actttgacgc cgtttattac tgactcgtcg aaagcttggc actggccgtc
10201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
10261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
10321 cagttgcgca gcctgaatgg cgaatgctag agcagcttga gcttggatca gattgtcgtt
10381 tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
10441 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt
10501 cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc
10561 caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa
10621 aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg
10681 cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga
10741 cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc
10801 gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg
10861 ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac
10921 cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc
10981 cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg
11041 gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc
11101 ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc
11161 gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg
11221 cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt
11281 ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc
11341 gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg
11401 gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg
11461 aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg
11521 agccgcccgc gcacgtctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg
11581 ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa
11641 aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc
11701 gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca
11761 gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc
11821 cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc
11881 cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga
11941 cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga
12001 cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc
12061 ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac
12121 ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg
12181 cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac
12241 gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc
12301 cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat
12361 ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt
12421 ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg
12481 aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta
12541 cgccaaggca agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa
12601 atgagcaaat gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa
12661 tcaagaacaa ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg
12721 gccaggcgta agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc
12781 cgaggaatcg gcgtgagcgg tcgcaaa
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pYPQ202植物表达质粒
别称: Plasmid#86198
| 原核抗性: | Kan |
|---|---|
| 筛选标记: | Hyg |
| 克隆菌株: | DB3.1 |
| 培养条件: | 37度 |
质粒属性
| 载体宿主： | 植物 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | |
| 基因类型： | ORF |
| 原核抗性： | Kan |
| 筛选标记： | Hyg |
| 荧光蛋白： |
质粒简介
pYPQ202是一个植物细胞Gateway系列表达质粒。
质粒图谱
质粒序列
LOCUS Exported 12807 bp ds-DNA circular SYN 12-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..12807
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1125..1319
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 1663..1803
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(1989..2577)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2664..3458)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 5082..5106
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 5184..5358
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(5399..6424)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(6491..7167)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
terminator 7489..7741
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 7805..7929
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(7970..8275)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLS-DK
VPRELYPVVHIGDESWRMMTTD-MASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(8595..9275)
/codon_start=1
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
DPAY"
promoter complement(9329..9359)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind complement(9384..9508)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature complement(9547..10181)
/label=AtUBQ10 promoter
misc_feature 10410..10434
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 11735..12364
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
1 ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag
61 gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg
121 caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg
181 tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat
241 gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag
301 cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt
361 tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt
421 tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc
481 gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag
541 cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag
601 cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt
661 agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta
721 gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac
781 cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc
841 gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc
901 gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg
961 ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac
1021 cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg
1081 caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac
1141 attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg
1201 tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt
1261 tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa
1321 ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc
1381 tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct
1441 ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc
1501 cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
1561 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
1621 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
1681 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
1741 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
1801 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
1861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
1921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
1981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
2041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
2101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
2161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
2221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
2281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
2341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
2401 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
2461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
2521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
2581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
2641 attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt
2701 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
2761 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
2821 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
2881 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
2941 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
3001 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
3061 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
3121 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
3181 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
3241 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
3301 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
3361 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
3421 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
3481 tacagtattt aaagataccc caagaagcta aggaaggtgc gaataagcgg ggaaattctt
3541 ctcggctgac tcagtcattt catttcttca tgtttgagcc gattttttct cccgtaaatg
3601 ccttgaatca gcctatttag accgtttctt cgccatttaa ggcgttatcc ccagttttta
3661 gtgagatctc tcccactgac gtatcatttg gtccgcccga aacaggttgg ccagcgtgaa
3721 taacatcgcc agttggttat cgtttttcag caaccccttg tatctggctt tcacgaagcc
3781 gaactgtcgc ttgatgatgc gaaatgggtg ctccaccctg gcccggatgc tggctttcat
3841 gtattcgatg ttgatggccg ttttgttctt gcgtggatgc tgtttcaagg ttcttacctt
3901 gccggggcgc tcggcgatca gccagtccac atccacctcg gccagctcct cgcgctgtgg
3961 cgccccttgg tagccggcat cggctgagac aaattgctcc tctccatgca gcagattacc
4021 cagctgattg aggtcatgct cgttggccgc ggtggtgacc aggctgtggg tcaggccact
4081 cttggcatcg acaccaatgt gggccttcat gccaaagtgc cactgattgc ctttcttggt
4141 ctgatgcatc tccggatcgc gttgctgctc tttgttcttg gtcgagctgg gtgcctcaat
4201 gatggtggca tcgaccaagg tgccttgagt catcatgacg cctgcttcgg ccagccagcg
4261 attgatggtc ttgaacaatt ggcgggccag ttgatgctgc tccagcaggt ggcggaaatt
4321 catgatggtg gtgcggtccg gcaaggcgct atccagggat aaccgggcaa acagacgcat
4381 ggaggcgatt tcgtacagag catcttccat cgcgccatcg ctcaggttgt accaatgctg
4441 catgcagtga atgcgtagca tggtttccag cggataaggt cgccggccat taccagcctt
4501 ggggtaaaac ggctcgatga cttccaccat gttttgccat ggcagaatct gctccatgcg
4561 ggacaagaaa atctcttttc tggtctgacg gcgcttactg ctgaattcac tgtcggcgaa
4621 ggtaagttga tgactcatga tgaaccctgt tctatggctc cagatgacaa acatgatctc
4681 atatcaggga cttgttcgca ccttccctaa ttataacaag acgaactcca attcactgtt
4741 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
4801 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
4861 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
4921 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
4981 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
5041 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
5101 gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactga
5161 attaacgccg aattaattcg ggggatctgg attttagtac tggattttgg ttttaggaat
5221 tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg gtttcttata
5281 tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg gaactactca
5341 cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg catctactct
5401 atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta
5461 cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc
5521 cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat
5581 tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga
5641 gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca
5701 tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga
5761 acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt
5821 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
5881 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
5941 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
6001 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
6061 tcgcatccat agcctccgcg accggttgta gaacagcggg cagttcggtt tcaggcaggt
6121 cttgcaacgt gacaccctgt gaacggcggg agatgcaata ggtcaggctc tcgctaaact
6181 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
6241 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
6301 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga
6361 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
6421 tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt gtagagagag
6481 actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt
6541 cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat
6601 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg
6661 ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat
6721 cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac
6781 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct
6841 caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt
6901 cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
6961 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
7021 tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt
7081 tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
7141 gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg
7201 atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg ctctagccaa
7261 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
7321 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
7381 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
7441 gataacaatt tcacacagga aacagctatg acatgattac gaattcccga tctagtaaca
7501 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
7561 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
7621 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
7681 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
7741 cggggaaatt cgagctccac cgcggtggcg gccgctctag aactagttaa ttaagaatta
7801 tcgaaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa atatcaatat
7861 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
7921 gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
7981 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
8041 cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
8101 catccccgat atgcaccacc gggtaaagtt cacgggggac tttatctgac agcagacgtg
8161 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
8221 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
8281 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
8341 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
8401 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
8461 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
8521 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
8581 ctgttatctg gcttttagta agccggatcc tctagattac gccccgcctg ccactcatcg
8641 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
8701 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
8761 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
8821 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
8881 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
8941 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
9001 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
9061 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
9121 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
9181 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
9241 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
9301 ctcgacggat cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag
9361 cctggggtgc ctaatgcggc cgccatagtg actggatatg ttgtgtttta cagtattatg
9421 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
9481 ctcgttcagc ttttttgtac aaacttgttt gatagcttgg cgcgcctcga gggggggccc
9541 ggtaccctgt taatcagaaa aactcagatt aatctacaaa ttcgatcgca caaactagaa
9601 actaacacca gatctagata gaaatcacaa atcgaagagt aattattcga caaaactcaa
9661 attatttgaa caaatcggat gatatctatg aaaccctaat cgagaattaa gatgatatct
9721 aacgatcaaa cccagaaaat cgtcttcgat ctaagattaa cagaatctaa accaaagaac
9781 atatacgaaa ttgggatcga acgaaaacaa aatcgaagat tttgagagaa taaggaacac
9841 agaaatttac cttgatcacg gtagagagaa ttgagagaaa gtttttaaga ttttgagaaa
9901 ttgaaatctg aattgtgaag aagaagagct ctttgggtat tgttttatag aagaagaaga
9961 agaaaagacg aggacgacta ggtcacgaga aagctaaggc ggtgaagcaa tagctaataa
10021 taaaatgaca cgtgtattga gcgttgttta cacgcaaagt tgtttttggc taattgcctt
10081 atttttaggt tgaggaaaag tatttgtgct ttgagttgat aaacacgact cgtgtgtgcc
10141 ggctgcaacc actttgacgc cgtttattac tgactcgtcg aaagcttggc actggccgtc
10201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
10261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
10321 cagttgcgca gcctgaatgg cgaatgctag agcagcttga gcttggatca gattgtcgtt
10381 tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
10441 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt
10501 cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc
10561 caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa
10621 aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg
10681 cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga
10741 cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc
10801 gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg
10861 ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac
10921 cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc
10981 cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg
11041 gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc
11101 ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc
11161 gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg
11221 cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt
11281 ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc
11341 gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg
11401 gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg
11461 aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg
11521 agccgcccgc gcacgtctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg
11581 ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa
11641 aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc
11701 gatgagtaaa taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca
11761 gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc
11821 cggggccgat gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc
11881 cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga
11941 cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga
12001 cttggctgtg tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc
12061 ttacgacata tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac
12121 ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg
12181 cggtgaggtt gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac
12241 gcagcgcgtg agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc
12301 cgagggcgac gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat
12361 ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt
12421 ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg
12481 aagcgggtca actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta
12541 cgccaaggca agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa
12601 atgagcaaat gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa
12661 tcaagaacaa ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg
12721 gccaggcgta agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc
12781 cgaggaatcg gcgtgagcgg tcgcaaa
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P3487/pENTR223-WBSCR28人源基因质粒 |
| P3488/pENTR223-AQP5人源基因质粒 |
| P3489/pENTR223-UNC50人源基因质粒 |
| P3491/pENTR223-NSA2人源基因质粒 |
| P3492/pENTR223-ECSIT人源基因质粒 |
| P3493/pENTR223-TSPAN5人源基因质粒 |
| P3494/pENTR223-CD27人源基因质粒 |
| P3495/pENTR223-KCNG4人源基因质粒 |
| P3496/pENTR223-SPIN1人源基因质粒 |
| P3497/pENTR223-RAB3IP人源基因质粒 |
| P3498/pENTR223-THAP12人源基因质粒 |
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文献和实验相关实验
ws 各位站友，本人因克隆某个植物抗病基因而完成了其BAC文库，目前正在做图位克隆的工作。 但文库的完成耗费了大量人力、物力和财力，因此想请教下该库还有什么其它用途。或与其 它技术结合，可开展什么工作。 谢谢了。。。。 zhujoker BAC (Bacterial Artificial Chromosome, 细菌人工染色体)文库是进行全基因组测序、构建物理图谱、染色体步
单子叶植物纲：包括11目，45科。 双子叶植物纲：2亚纲，44目，259科。 原始花被亚纲（离瓣花亚纲）：33目，202科。 后生花被亚纲（合瓣花亚纲）：11目，57科。 恩氏系统是根据假花说的原理，认为无瓣花、单性化、风媒传粉、木本等性状是原始的特征；认为有花瓣、两性花、虫媒传粉是进化的特征。为此把柔荑花序类植物（胡桃科、壳斗科、木麻黄科…）当作被子植物中最原始类型，而把木兰科、毛茛科等看作是较进化的类型。1964
鸦雀属 鸦雀属 （Paradoxornis；crowtits，parrotbills） 雀形目鹟科画眉亚科的1属。体型较小，体长90～202毫米。嘴形短厚，其厚度较长度为大，或与长度几乎相等；鼻孔完全被羽掩盖；翅短而圆，第一枚初级飞羽的长度超过第二枚之半。翅长不及100毫米。世界共有17种，中国有14种，棕头鸦雀（见图）为本属的常见种，遍布中国北部、东部和南部。头至上背红棕色，上体余部为带黄的橄榄褐色；两翅和尾羽暗褐色，颏、喉和上胸玫瑰棕色；腹部中央皮黄色。常集
pYPQ202植物表达质粒
￥100 - 1000
