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- 详细信息
- 询价记录
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
pDG1730
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
质粒干粉
基本信息
名称：pDG1730枯草胞内质粒
别称: pDG1730
质粒属性
质粒简介
pDG1730质粒由上海烜雅生物科技有限公司提供
质粒图谱
质粒序列
LOCUS Exported 7602 bp ds-DNA circular SYN 27-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7602)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7602
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 115..693
/codon_start=1
/label=amyE-front
/translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP
SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ
PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH
GNTQIKNWSDRILEAYRIPAALANGDFRS"
CDS 1039..1713
/codon_start=1
/label=Spe
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
CDS 1849..2868
/codon_start=1
/label=amyE-back
/translation="MSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQIGDR
GSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSVSIN
TATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKTGVT
HSFNDQLTITLRADANTTKAVYQINNGPDDRRLRMEINSQSEKEIQFGKTYTIMLKGTN
SDGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGSRVIELTGSWPGKPMT
KNADGIYTLTLPADTDTTNAKVIFNNGSAQVPGQNQPGFDYVLNGLYNDSGLSGSLPH"
CDS 4231..4965
/codon_start=1
/label=Erm
/translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
DLNNISFEQFLSLFNSYKLFNK"
rep_origin complement(5773..6361)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6532..7392)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ORIGIN
1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
601 catggaaaca cacaaattaa aaactggtct gatcggatcc tagaagctta tcgaattcct
661 gcagccctgg cgaatggcga ttttcgttcg tgaatacatg ttataataac tataactaat
721 aacgtaacgt gactggcaag agatattttt aaaacaatga ataggtttac acttacttta
781 gttttatgga aatgaaagat catatcatat ataatctaga ataaaattaa ctaaaataat
841 tattatctag ataaaaaatt tagaagccaa tgaaatctat aaataaacta aattaagttt
901 atttaattaa caactatgga tataaaatag gtactaatca aaatagtgag gaggatatat
961 ttgaatacat acgaacaaat taataaagtg aaaaaaatac ttcggaaaca tttaaaaaat
1021 aaccttattg gtacttacat gtttggatca ggagttgaga gtggactaaa accaaatagt
1081 gatcttgact ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa agaaatactt
1141 atacaaaaaa ttagacctat ttcaaaaaaa ataggagata aaagcaactt acgatatatt
1201 gaattaacaa ttattattca gcaagaaatg gtaccgtgga atcatcctcc caaacaagaa
1261 tttatttatg gagaatggtt acaagagctt tatgaacaag gatacattcc tcagaaggaa
1321 ttaaattcag atttaaccat aatgctttac caagcaaaac gaaaaaataa aagaatatac
1381 ggaaattatg acttagagga attactacct gatattccat tttctgatgt gagaagagcc
1441 attatggatt cgtcagagga attaatagat aattatcagg atgatgaaac caactctata
1501 ttaactttat gccgtatgat tttaactatg gacacgggta aaatcatacc aaaagatatt
1561 gcgggaaatg cagtggctga atcttctcca ttagaacata gggagagaat tttgttagca
1621 gttcgtagtt atcttggaga gaatattgaa tggactaatg aaaatgtaaa tttaactata
1681 aactatttaa ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg tggaaacact
1741 tttttcaatt tttttgtttt attatttaat atttgggaaa tattcattct aattggtaat
1801 cagattttag aaaacaataa acccttgcat agggggatct cgacatggat gagcgatgat
1861 gatatccgtt taggctgggc ggtgatagct tctcgttcag gcagtacgcc tcttttcttt
1921 tccagacctg agggaggcgg aaatggtgtg aggttcccgg ggaaaagcca aataggcgat
1981 cgcgggagtg ctttatttga agatcaggct atcactgcgg tcaatagatt tcacaatgtg
2041 atggctggac agcctgagga actctcgaac ccgaatggaa acaaccagat atttatgaat
2101 cagcgcggct cacatggcgt tgtgctggca aatgcaggtt catcctctgt ctctatcaat
2161 acggcaacaa aattgcctga tggcaggtat gacaataaag ctggagcggg ttcatttcaa
2221 gtgaacgatg gtaaactgac aggcacgatc aatgccaggt ctgtagctgt gctttatcct
2281 gatgatattg caaaagcgcc tcatgttttc cttgagaatt acaaaacagg tgtaacacat
2341 tctttcaatg atcaactgac gattaccttg cgtgcagatg cgaatacaac aaaagccgtt
2401 tatcaaatca ataatggacc agacgacagg cgtttaagga tggagatcaa ttcacaatcg
2461 gaaaaggaga tccaatttgg caaaacatac accatcatgt taaaaggaac gaacagtgat
2521 ggtgtaacga ggaccgagaa atacagtttt gttaaaagag atccagcgtc ggccaaaacc
2581 atcggctatc aaaatccgaa tcattggagc caggtaaatg cttatatcta taaacatgat
2641 gggagccgag taattgaatt gaccggatct tggcctggaa aaccaatgac taaaaatgca
2701 gacggaattt acacgctgac gctgcctgcg gacacggata caaccaacgc aaaagtgatt
2761 tttaataatg gcagcgccca agtgcccggt cagaatcagc ctggctttga ttacgtgcta
2821 aatggtttat ataatgactc gggcttaagc ggttctcttc cccattgagg gcaaggctag
2881 acgggactta ccgaaagaaa ccatcaatga tggtttcttt tttgttcata aatcagacaa
2941 aacttttctc ttgcaaaagt ttgtgaagtg ttgcacaata taaatgtgaa atacttcaca
3001 aacaaaaaga catcaaagag aaacataccc tgcaaggatg ctgatattgt ctgcatttgc
3061 gccggagcaa accaaaaacc tggtgagaca cgccttgaat tagtagaaaa gaacttgaag
3121 attttcaaag gcatcgttag tgaagtcatg gcgagcggat ttgacggcat tttcttagtc
3181 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
3241 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
3301 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg
3361 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
3421 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctactga
3481 agtggatttc tttaagagct cctttaactt cctcaccagt agttgtatcg gtaccataag
3541 tagaagcagc aacccaagta gctttaccag catccggttc aaccagcata gtaagaatct
3601 tactggacat cggcagttct tcgaacagtg cgccaactac cagctctttc tgcagttcat
3661 tcagggcacc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
3721 tcctgtctct tgatccatgg attacgcgtt aacccgggcc cgcggatgca tatgatcaga
3781 tcctttaact ctggcaaccc tcaaaattga atgagacatg ctacacctcc ggataataaa
3841 tatatataaa cgtatataga tttcataaag tctaacacac tagacttatt tacttcgtaa
3901 ttaagtcgtt aaaccgtgtg ctctacgacc aaaactataa aacctttaag aactttcttt
3961 ttttacaaga aaaaagaaat tagataaatc tctcatatct tttattcaat aatcgcatcc
4021 gattgcagta taaatttaac gatcactcat catgttcata tttatcagag ctcgtgctat
4081 aattatacta attttataag gaggaaaaaa tatgggcatt tttagtattt ttgtaatcag
4141 cacagttcat tatcaaccaa acaaaaaata agtggttata atgaatcgtt aataagcaaa
4201 attcatataa ccaaattaaa gagggttata atgaacgaga aaaatataaa acacagtcaa
4261 aactttatta cttcaaaaca taatatagat aaaataatga caaatataag attaaatgaa
4321 catgataata tctttgaaat cggctcagga aaaggccatt ttacccttga attagtaaag
4381 aggtgtaatt tcgtaactgc cattgaaata gaccataaat tatgcaaaac tacagaaaat
4441 aaacttgttg atcacgataa tttccaagtt ttaaacaagg atatattgca gtttaaattt
4501 cctaaaaacc aatcctataa aatatatggt aatatacctt ataacataag tacggatata
4561 atacgcaaaa ttgtttttga tagtatagct aatgagattt atttaatcgt ggaatacggg
4621 tttgctaaaa gattattaaa tacaaaacgc tcattggcat tacttttaat ggcagaagtt
4681 gatatttcta tattaagtat ggttccaaga gaatattttc atcctaaacc taaagtgaat
4741 agctcactta tcagattaag tagaaaaaaa tcaagaatat cacacaaaga taaacaaaag
4801 tataattatt tcgttatgaa atgggttaac aaagaataca agaaaatatt tacaaaaaat
4861 caatttaaca attccttaaa acatgcagga attgacgatt taaacaatat tagctttgaa
4921 caattcttat ctcttttcaa tagctataaa ttatttaata agtaagttaa gggatgcata
4981 aactgcatcc cttaacttgt ttttcgtgtg cctatttttt gtgaatcgat tatgtctttt
5041 gcgcagtcgg cttaaaccag ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa
5101 attcaaaatc tatcggtcag atttataccg atttgatttt atatattctt gaataacata
5161 cgccgagtta tcacataaaa gcgggaacca atcatcaaat ttaaacttca ttgcataatc
5221 cattaaactc ttaaattcta cgattccttg ttcatcaata aactcaatca tttctttaat
5281 taatttatat ctatctgttg ttgttttctt taataattca tcaacatcta caccgccata
5341 aactatcata tcttcttttt gatatttaaa tttattagga tcttaaggcc taggtctaga
5401 gtctttgttt tgacgccatt agcgtacgta acaatcctcg ttaaaggaca aggacctgag
5461 cggaagtgta tcgtacagta gacggagtat actagtatag tctatagtcc gtggaattat
5521 tatatttatc tccgacgata ttctcatcag tgaaatccag ctggagttct ttagcaaatt
5581 tttttattag ctgaacttag tattagtggc catactcctc caatccaaag ctatttagaa
5641 agattactat atcctcaaac aggcggtaac cggcctcttc atcgggaatg cgcgcgacct
5701 tcagcatcgc cggcatgtcc ccctggcgga cgggaagtat ccagctcgag gtcgggccgc
5761 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
5821 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
5881 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
5941 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
6001 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
6061 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
6121 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
6181 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
6241 gaagccagtt accttcggaa aaagagttga tagctcttga tccggcaaac aaaccaccgc
6301 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
6361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
6421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
6481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
6541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
6601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
6661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
6721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
6781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
6841 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
6901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
6961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
7021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
7081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
7141 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
7201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
7261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
7321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
7381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
7441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
7501 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
7561 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa tt
//
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：pDG1730枯草胞内质粒
别称: pDG1730
| 原核抗性: | Amp |
|---|---|
| 筛选标记: | Erm,Spe |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37度 |
质粒属性
| 载体宿主： | 枯草杆菌 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： |
质粒简介
pDG1730质粒由上海烜雅生物科技有限公司提供
质粒图谱
质粒序列
LOCUS Exported 7602 bp ds-DNA circular SYN 27-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7602)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7602
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 115..693
/codon_start=1
/label=amyE-front
/translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP
SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ
PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH
GNTQIKNWSDRILEAYRIPAALANGDFRS"
CDS 1039..1713
/codon_start=1
/label=Spe
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
CDS 1849..2868
/codon_start=1
/label=amyE-back
/translation="MSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQIGDR
GSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSVSIN
TATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKTGVT
HSFNDQLTITLRADANTTKAVYQINNGPDDRRLRMEINSQSEKEIQFGKTYTIMLKGTN
SDGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGSRVIELTGSWPGKPMT
KNADGIYTLTLPADTDTTNAKVIFNNGSAQVPGQNQPGFDYVLNGLYNDSGLSGSLPH"
CDS 4231..4965
/codon_start=1
/label=Erm
/translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
DLNNISFEQFLSLFNSYKLFNK"
rep_origin complement(5773..6361)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6532..7392)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ORIGIN
1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
601 catggaaaca cacaaattaa aaactggtct gatcggatcc tagaagctta tcgaattcct
661 gcagccctgg cgaatggcga ttttcgttcg tgaatacatg ttataataac tataactaat
721 aacgtaacgt gactggcaag agatattttt aaaacaatga ataggtttac acttacttta
781 gttttatgga aatgaaagat catatcatat ataatctaga ataaaattaa ctaaaataat
841 tattatctag ataaaaaatt tagaagccaa tgaaatctat aaataaacta aattaagttt
901 atttaattaa caactatgga tataaaatag gtactaatca aaatagtgag gaggatatat
961 ttgaatacat acgaacaaat taataaagtg aaaaaaatac ttcggaaaca tttaaaaaat
1021 aaccttattg gtacttacat gtttggatca ggagttgaga gtggactaaa accaaatagt
1081 gatcttgact ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa agaaatactt
1141 atacaaaaaa ttagacctat ttcaaaaaaa ataggagata aaagcaactt acgatatatt
1201 gaattaacaa ttattattca gcaagaaatg gtaccgtgga atcatcctcc caaacaagaa
1261 tttatttatg gagaatggtt acaagagctt tatgaacaag gatacattcc tcagaaggaa
1321 ttaaattcag atttaaccat aatgctttac caagcaaaac gaaaaaataa aagaatatac
1381 ggaaattatg acttagagga attactacct gatattccat tttctgatgt gagaagagcc
1441 attatggatt cgtcagagga attaatagat aattatcagg atgatgaaac caactctata
1501 ttaactttat gccgtatgat tttaactatg gacacgggta aaatcatacc aaaagatatt
1561 gcgggaaatg cagtggctga atcttctcca ttagaacata gggagagaat tttgttagca
1621 gttcgtagtt atcttggaga gaatattgaa tggactaatg aaaatgtaaa tttaactata
1681 aactatttaa ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg tggaaacact
1741 tttttcaatt tttttgtttt attatttaat atttgggaaa tattcattct aattggtaat
1801 cagattttag aaaacaataa acccttgcat agggggatct cgacatggat gagcgatgat
1861 gatatccgtt taggctgggc ggtgatagct tctcgttcag gcagtacgcc tcttttcttt
1921 tccagacctg agggaggcgg aaatggtgtg aggttcccgg ggaaaagcca aataggcgat
1981 cgcgggagtg ctttatttga agatcaggct atcactgcgg tcaatagatt tcacaatgtg
2041 atggctggac agcctgagga actctcgaac ccgaatggaa acaaccagat atttatgaat
2101 cagcgcggct cacatggcgt tgtgctggca aatgcaggtt catcctctgt ctctatcaat
2161 acggcaacaa aattgcctga tggcaggtat gacaataaag ctggagcggg ttcatttcaa
2221 gtgaacgatg gtaaactgac aggcacgatc aatgccaggt ctgtagctgt gctttatcct
2281 gatgatattg caaaagcgcc tcatgttttc cttgagaatt acaaaacagg tgtaacacat
2341 tctttcaatg atcaactgac gattaccttg cgtgcagatg cgaatacaac aaaagccgtt
2401 tatcaaatca ataatggacc agacgacagg cgtttaagga tggagatcaa ttcacaatcg
2461 gaaaaggaga tccaatttgg caaaacatac accatcatgt taaaaggaac gaacagtgat
2521 ggtgtaacga ggaccgagaa atacagtttt gttaaaagag atccagcgtc ggccaaaacc
2581 atcggctatc aaaatccgaa tcattggagc caggtaaatg cttatatcta taaacatgat
2641 gggagccgag taattgaatt gaccggatct tggcctggaa aaccaatgac taaaaatgca
2701 gacggaattt acacgctgac gctgcctgcg gacacggata caaccaacgc aaaagtgatt
2761 tttaataatg gcagcgccca agtgcccggt cagaatcagc ctggctttga ttacgtgcta
2821 aatggtttat ataatgactc gggcttaagc ggttctcttc cccattgagg gcaaggctag
2881 acgggactta ccgaaagaaa ccatcaatga tggtttcttt tttgttcata aatcagacaa
2941 aacttttctc ttgcaaaagt ttgtgaagtg ttgcacaata taaatgtgaa atacttcaca
3001 aacaaaaaga catcaaagag aaacataccc tgcaaggatg ctgatattgt ctgcatttgc
3061 gccggagcaa accaaaaacc tggtgagaca cgccttgaat tagtagaaaa gaacttgaag
3121 attttcaaag gcatcgttag tgaagtcatg gcgagcggat ttgacggcat tttcttagtc
3181 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
3241 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
3301 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg
3361 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
3421 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctactga
3481 agtggatttc tttaagagct cctttaactt cctcaccagt agttgtatcg gtaccataag
3541 tagaagcagc aacccaagta gctttaccag catccggttc aaccagcata gtaagaatct
3601 tactggacat cggcagttct tcgaacagtg cgccaactac cagctctttc tgcagttcat
3661 tcagggcacc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
3721 tcctgtctct tgatccatgg attacgcgtt aacccgggcc cgcggatgca tatgatcaga
3781 tcctttaact ctggcaaccc tcaaaattga atgagacatg ctacacctcc ggataataaa
3841 tatatataaa cgtatataga tttcataaag tctaacacac tagacttatt tacttcgtaa
3901 ttaagtcgtt aaaccgtgtg ctctacgacc aaaactataa aacctttaag aactttcttt
3961 ttttacaaga aaaaagaaat tagataaatc tctcatatct tttattcaat aatcgcatcc
4021 gattgcagta taaatttaac gatcactcat catgttcata tttatcagag ctcgtgctat
4081 aattatacta attttataag gaggaaaaaa tatgggcatt tttagtattt ttgtaatcag
4141 cacagttcat tatcaaccaa acaaaaaata agtggttata atgaatcgtt aataagcaaa
4201 attcatataa ccaaattaaa gagggttata atgaacgaga aaaatataaa acacagtcaa
4261 aactttatta cttcaaaaca taatatagat aaaataatga caaatataag attaaatgaa
4321 catgataata tctttgaaat cggctcagga aaaggccatt ttacccttga attagtaaag
4381 aggtgtaatt tcgtaactgc cattgaaata gaccataaat tatgcaaaac tacagaaaat
4441 aaacttgttg atcacgataa tttccaagtt ttaaacaagg atatattgca gtttaaattt
4501 cctaaaaacc aatcctataa aatatatggt aatatacctt ataacataag tacggatata
4561 atacgcaaaa ttgtttttga tagtatagct aatgagattt atttaatcgt ggaatacggg
4621 tttgctaaaa gattattaaa tacaaaacgc tcattggcat tacttttaat ggcagaagtt
4681 gatatttcta tattaagtat ggttccaaga gaatattttc atcctaaacc taaagtgaat
4741 agctcactta tcagattaag tagaaaaaaa tcaagaatat cacacaaaga taaacaaaag
4801 tataattatt tcgttatgaa atgggttaac aaagaataca agaaaatatt tacaaaaaat
4861 caatttaaca attccttaaa acatgcagga attgacgatt taaacaatat tagctttgaa
4921 caattcttat ctcttttcaa tagctataaa ttatttaata agtaagttaa gggatgcata
4981 aactgcatcc cttaacttgt ttttcgtgtg cctatttttt gtgaatcgat tatgtctttt
5041 gcgcagtcgg cttaaaccag ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa
5101 attcaaaatc tatcggtcag atttataccg atttgatttt atatattctt gaataacata
5161 cgccgagtta tcacataaaa gcgggaacca atcatcaaat ttaaacttca ttgcataatc
5221 cattaaactc ttaaattcta cgattccttg ttcatcaata aactcaatca tttctttaat
5281 taatttatat ctatctgttg ttgttttctt taataattca tcaacatcta caccgccata
5341 aactatcata tcttcttttt gatatttaaa tttattagga tcttaaggcc taggtctaga
5401 gtctttgttt tgacgccatt agcgtacgta acaatcctcg ttaaaggaca aggacctgag
5461 cggaagtgta tcgtacagta gacggagtat actagtatag tctatagtcc gtggaattat
5521 tatatttatc tccgacgata ttctcatcag tgaaatccag ctggagttct ttagcaaatt
5581 tttttattag ctgaacttag tattagtggc catactcctc caatccaaag ctatttagaa
5641 agattactat atcctcaaac aggcggtaac cggcctcttc atcgggaatg cgcgcgacct
5701 tcagcatcgc cggcatgtcc ccctggcgga cgggaagtat ccagctcgag gtcgggccgc
5761 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
5821 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
5881 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
5941 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
6001 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
6061 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
6121 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
6181 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
6241 gaagccagtt accttcggaa aaagagttga tagctcttga tccggcaaac aaaccaccgc
6301 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
6361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
6421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
6481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
6541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
6601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
6661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
6721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
6781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
6841 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
6901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
6961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
7021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
7081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
7141 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
7201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
7261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
7321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
7381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
7441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
7501 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
7561 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa tt
//
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
| P4444/pCMV-SPORT6-NRL人源基因质粒 |
| P4445/pCMV-SPORT6-CAVIN2人源基因质粒 |
| P4446/pCMV-SPORT6-SQOR人源基因质粒 |
| P4447/pCMV-SPORT6-TOX4（1同义突变）人源基因质粒 |
| P4448/pCMV-SPORT6-E2F3人源基因质粒 |
| P4449/pCMV-SPORT6-ANGPTL2人源基因质粒 |
| P4450/pCMV-SPORT6-VDAC2人源基因质粒 |
| P4451/pCMV-SPORT6-SRGN（1点突变）人源基因质粒 |
| P4452/pCMV-SPORT6-ALKBH3人源基因质粒 |
| P4453/pCMV-SPORT6-PEX11B（1点突变）人源基因质粒 |
| P4454/pCMV-SPORT6-CLDND1人源基因质粒 |
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pDG1730枯草胞内质粒
￥1280
