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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Super PiggyBac Transposase
- 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：Super PiggyBac Transposase (PB200PA-1) 哺乳转座质粒
别称: Super PiggyBac Transposase
质粒属性
质粒简介
Super PiggyBac Transposase (PB200PA-1)是一个哺乳细胞转座质粒。The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors andchromosomes via a "cut and paste" mechanism. During transposition, the PB transposase recognizes transposonspecific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes.The Super PiggyBac transposase expression vector features a 5’ HS4 insulator to support robust transcription from the rPolr2A promoter. The PiggyBac transposase coding sequence has been optimized for high expression, stability and activity in mammalian cells.
质粒图谱
质粒序列
LOCUS Exported 8531 bp ds-DNA circular SYN 17-AUG-2018
DEFINITION synthetic circular DNA
KEYWORDS Super PiggyBac Transposase(PB200PA-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8531)
TITLE Direct Submission
JOURNAL Exported Friday, August 17, 2018 from SnapGene 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8531
/organism="synthetic DNA construct"
/mol_type="other DNA"
intron 3442..3538
/note="SV40 intron"
/note="modified SV40 intron with splice donor and acceptor
sites"
regulatory 3608..3617
/regulatory_class="other"
/note="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3614..5398
/codon_start=1
/note="Super PiggyBac Transposase"
/translation="MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEE
AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKST
RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
TSATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
CLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFR
VYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSC
RNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDG
PLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTC
SRKTNRWP-MALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRK
RLEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKK
CKKVICREHNIDMCQSCF"
polyA_signal 5424..5558
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(6116..6704)
/direction=LEFT
/note="pUC ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6875..7735)
/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(7736..7840)
/gene="bla"
/note="AmpR promoter"
rep_origin 7867..8322
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 gcggccgcga gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
61 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
121 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
181 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
241 gggaaaaagc tttaggctga aagagagatt tagaatgaca gaatcataga acggcctggg
301 ttgcaaagga gcacagtgct catccagatc caaccccctg ctatgtgcag ggtcatcaac
361 cagcagccca ggctgcccag agccacatcc agcctggcct tgaatgcctg cagggatggg
421 gcatccacag cctccttggg caacctgttc agtgcgtcac caccctctgg gggaaaaact
481 gcctcctcat atccaaccca aacctcccct gtctcagtgt aaagccattc ccccttgtcc
541 tatcaagggg gagtttgctg tgacattgtt ggtctggggt gacacatgtt tgccaattca
601 gtgcatcacg gagaggcaga tcttggggat aaggaagtgc aggacagcat ggacgtggga
661 catgcaggtg ttgagggctc tgggacactc tccaagtcac agcgttcaga acagccttaa
721 ggataagaag ataggataga aggacaaaga gcaagttaaa acccagcatg gagaggagca
781 caaaaaggcc acagacactg ctggtccctg tgtctgagcc tgcatgtttg atggtgtctg
841 gatgcaagca gaaggggtgg aagagcttgc ctggagagat acagctgggt cagtaggact
901 gggacaggca gctggagaat tgccatgtag atgttcatac aatcgtcaaa tcatgaaggc
961 tggaaaagcc ctccaagatc cccaagacca accccaaccc acccaccgtg cccactggcc
1021 atgtccctca gtgccacatc cccacagttc ttcatcacct ccagggacgg tgaccccccc
1081 acctccgtgg gcagctgtgc cactgcagca ccgctctttg gagaaggtaa atcttgctaa
1141 atccagcccg accctcccct ggcacaacgt aaggccatta tctctcatcc aactccagga
1201 cggagtcagt gagaatattg cctccctggc gagctcacgg ggacagcccc cccccaaagc
1261 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
1321 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
1381 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
1441 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
1501 cagaatcata gaacggcctg ggttgcaaag gagcacagtg ctcatccaga tccaaccccc
1561 tgctatgtgc agggtcatca accagcagcc caggctgccc agagccacat ccagcctggc
1621 cttgaatgcc tgcagggatg gggcatccac agcctccttg ggcaacctgt tcagtgcgtc
1681 accaccctct gggggaaaaa ctgcctcctc atatccaacc caaacctccc ctgtctcagt
1741 gtaaagccat tcccccttgt cctatcaagg gggagtttgc tgtgacattg ttggtctggg
1801 gtgacacatg tttgccaatt cagtgcatca cggagaggca gatcttgggg ataaggaagt
1861 gcaggacagc atggacgtgg gacatgcagg tgttgagggc tctgggacac tctccaagtc
1921 acagcgttca gaacagcctt aaggataaga agataggata gaaggacaaa gagcaagtta
1981 aaacccagca tggagaggag cacaaaaagg ccacagacac tgctggtccc tgtgtctgag
2041 cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt ggaagagctt gcctggagag
2101 atacagctgg gtcagtagga ctgggacagg cagctggaga attgccatgt agatgttcat
2161 acaatcgtca aatcatgaag gctggaaaag ccctccaaga tccccaagac caaccccaac
2221 ccacccaccg tgcccactgg ccatgtccct cagtgccaca tccccacagt tcttcatcac
2281 ctccagggac ggtgaccccc ccacctccgt gggcagctgt gccactgcag caccgctctt
2341 tggagaaggt aaatcttgct aaatccagcc cgaccctccc ctggcacaac gtaaggccat
2401 tatctctcat ccaactccag gacggagtca gtgagaatat tcggccgctc tagaactagt
2461 ggatcccccg ggctgcagga attcgatggc cgcttgggcg ggatatctag cgccaaggtt
2521 ccgtaaagct ctctattaga ggactggcgg tgccccatca ccctgggagg cctctccaca
2581 aatatcggct acttccaatt gattggacgc gccatcttgt ctgcttatgc atattcagag
2641 gatcctgaat attcatgagc gagggccgtg cggcccctcc ctccaaccct cccccggaac
2701 aagctccgga gaacccgaca ggccccgcct tctttaccga tgcgtagaac aaaccatttt
2761 ccgggttggg gggggaaatt aatgagagac tttagctgaa aatgagcatg gacgccaaag
2821 ctgagtaaag atggcttaac tttatcctcc attctgtaat ccgtcagctt gagtgtacgg
2881 gaagtcggcg aggggggcgg caggctcaga aacattctcc tcctcctgtc gcgtcagaaa
2941 gaacacccaa ccagggagcc ggagccctgg cgtcaacatc tctggcgcgc gcgctccatg
3001 taggccggtg cgggcggccc cgtagcgcaa gggagggcgg gaaaggaagg ggcgggacac
3061 aagggcgaat ctataaaggg cgtcattcag ccagttctct cctcagaagc gccgagagcg
3121 cgaccgggac ggttggagaa gaaggtggct cccggaaggg ggagagacaa actgccgtaa
3181 cctctgccgt tcaggatccc ggccatcaag cttatcgatg ctgccgcagc aaaagcagga
3241 gcagatgccg ccgtcgcagg cgaagatgtc gcagacggag gaggcgatgc tgccggcgga
3301 ggaggcgaag taagtagagg gctgggctgg gctgtggggg gtgtggggtg cgggactggg
3361 cagtctggga gtccctctca ccacttttct tacctttcta ggatgctgcc tcgaggaact
3421 gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga
3481 tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc
3541 ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat tgtacccaat tcgttaaggc
3601 caaattggcc accatgggct ctagcctgga cgacgagcac atcctgagcg ccctgctgca
3661 gagcgacgac gaactggtgg gcgaggacag cgacagcgag gtcagcgacc acgtgtccga
3721 ggacgacgtg cagtccgaca ccgaggaagc cttcatcgac gaggtgcacg aagtgcagcc
3781 taccagcagc ggctccgaga tcctggacga gcagaacgtg atcgagcagc ctggcagctc
3841 cctggccagc aacagaatcc tgaccctgcc ccagagaacc atcagaggca agaacaagca
3901 ctgctggtcc acctccaaga gcaccaggcg gagcagagtg tccgccctga acatcgtgcg
3961 gagccagagg ggccccacca gaatgtgcag aaacatctac gaccccctgc tgtgcttcaa
4021 gctgttcttc accgacgaga tcatcagcga gatcgtgaag tggaccaacg ccgagatcag
4081 cctgaagagg cgggagagca tgaccagcgc caccttcaga gacaccaacg aggacgagat
4141 ctacgccttc ttcggcatcc tggtgatgac cgccgtgaga aaggacaacc acatgagcac
4201 cgacgacctg ttcgacagat ccctgagcat ggtgtacgtg tccgtgatga gcagagacag
4261 attcgacttc ctgatcagat gcctgagaat ggacgacaag agcatcagac ccaccctgcg
4321 ggagaacgac gtgttcaccc ccgtgcggaa gatctgggac ctgttcatcc accagtgcat
4381 ccagaactac acccctggcg cccacctgac catcgatgag cagctgctgg gcttcagagg
4441 cagatgcccc ttcagagtgt acatccccaa caagcccagc aagtacggca tcaagatcct
4501 gatgatgtgc gacagcggca ccaagtacat gatcaacggc atgccctacc tgggcagagg
4561 cacccagaca aacggcgtgc ccctgggcga gtactacgtg aaagaactga gcaagcctgt
4621 gcatggcagc tgcaggaaca tcacctgcga caactggttc accagcatcc ccctggccaa
4681 gaacctgctg caggaaccct acaagctgac catcgtgggc accgtgcgga gcaacaagcg
4741 ggagatccca gaggtgctga agaacagcag atccagacct gtgggaacaa gcatgttctg
4801 cttcgacggc cccctgaccc tggtgtccta caagcccaag cccgccaaga tggtgtacct
4861 gctgtccagc tgcgacgagg acgccagcat caacgagagc accggcaagc cccagatggt
4921 gatgtactac aaccagacca agggcggcgt ggacaccctg gaccagatgt gcagcgtgat
4981 gacctgcagc agaaagacca acagatggcc catggccctg ctgtacggca tgatcaatat
5041 cgcctgcatc aacagcttca tcatctacag ccacaacgtg tccagcaagg gcgagaaggt
5101 gcagagccgg aagaaattca tgcggaacct gtacatgagc ctgacctcca gcttcatgag
5161 aaagagactg gaagccccca ccctgaagag atacctgcgg gacaacatca gcaacatcct
5221 gcccaaggaa gtgccaggaa caagcgacga cagcaccgag gaacccgtga tgaagaagag
5281 gacctactgc acctactgtc ccagcaagat cagaagaaag gccaacgcca gctgcaagaa
5341 atgcaaaaaa gtgatctgcc gggagcacaa catcgacatg tgccagagct gtttctgagg
5401 ccgtaacggc cgccagaatt ggggatccag acatgataag atacattgat gagtttggac
5461 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
5521 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
5581 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgacct
5641 gcaggcatgc aagctcggta cccagctttt gttcccttta gtgagggtta attgcgcgct
5701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
5761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
5821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
5881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
5941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
6001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
6061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
6121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
6181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
6241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
6301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
6361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
6421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
6481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
6541 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
6601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
6661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
6721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
6781 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
6841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
6901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
6961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
7021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
7081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
7141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
7201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
7261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
7321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
7381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
7441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
7501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
7561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
7621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
7681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
7741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
7801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
7861 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
7921 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
7981 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
8041 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
8101 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
8161 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
8221 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
8281 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
8341 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
8401 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
8461 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct
8521 ccaccgcggt g
//
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：Super PiggyBac Transposase (PB200PA-1) 哺乳转座质粒
别称: Super PiggyBac Transposase
| 启动子: | Polr2A |
|---|---|
| 复制子: | pUC |
| 终止子: | SV40 poly(A) signal |
| 质粒分类: | 哺乳细胞转座酶表达质粒 |
| 质粒大小: | 8531bp |
| 原核抗性: | Amp |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37度 |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 无须诱导，瞬时表达 |
| 5'测序引物: | M13R:CAGGAAACAGCTATGACC |
| 3'测序引物: | 根据序列设计引物 |
质粒属性
| 载体宿主： | 哺乳细胞 |
|---|---|
| 载体用途： | 蛋白表达,转座 |
| 基因种属： | |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 真核抗性： | |
| 荧光蛋白： |
质粒简介
Super PiggyBac Transposase (PB200PA-1)是一个哺乳细胞转座质粒。The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors andchromosomes via a "cut and paste" mechanism. During transposition, the PB transposase recognizes transposonspecific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes.The Super PiggyBac transposase expression vector features a 5’ HS4 insulator to support robust transcription from the rPolr2A promoter. The PiggyBac transposase coding sequence has been optimized for high expression, stability and activity in mammalian cells.
质粒图谱
质粒序列
LOCUS Exported 8531 bp ds-DNA circular SYN 17-AUG-2018
DEFINITION synthetic circular DNA
KEYWORDS Super PiggyBac Transposase(PB200PA-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8531)
TITLE Direct Submission
JOURNAL Exported Friday, August 17, 2018 from SnapGene 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8531
/organism="synthetic DNA construct"
/mol_type="other DNA"
intron 3442..3538
/note="SV40 intron"
/note="modified SV40 intron with splice donor and acceptor
sites"
regulatory 3608..3617
/regulatory_class="other"
/note="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3614..5398
/codon_start=1
/note="Super PiggyBac Transposase"
/translation="MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEE
AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKST
RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
TSATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
CLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFR
VYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSC
RNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDG
PLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTC
SRKTNRWP-MALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRK
RLEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKK
CKKVICREHNIDMCQSCF"
polyA_signal 5424..5558
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(6116..6704)
/direction=LEFT
/note="pUC ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6875..7735)
/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(7736..7840)
/gene="bla"
/note="AmpR promoter"
rep_origin 7867..8322
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 gcggccgcga gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
61 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
121 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
181 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
241 gggaaaaagc tttaggctga aagagagatt tagaatgaca gaatcataga acggcctggg
301 ttgcaaagga gcacagtgct catccagatc caaccccctg ctatgtgcag ggtcatcaac
361 cagcagccca ggctgcccag agccacatcc agcctggcct tgaatgcctg cagggatggg
421 gcatccacag cctccttggg caacctgttc agtgcgtcac caccctctgg gggaaaaact
481 gcctcctcat atccaaccca aacctcccct gtctcagtgt aaagccattc ccccttgtcc
541 tatcaagggg gagtttgctg tgacattgtt ggtctggggt gacacatgtt tgccaattca
601 gtgcatcacg gagaggcaga tcttggggat aaggaagtgc aggacagcat ggacgtggga
661 catgcaggtg ttgagggctc tgggacactc tccaagtcac agcgttcaga acagccttaa
721 ggataagaag ataggataga aggacaaaga gcaagttaaa acccagcatg gagaggagca
781 caaaaaggcc acagacactg ctggtccctg tgtctgagcc tgcatgtttg atggtgtctg
841 gatgcaagca gaaggggtgg aagagcttgc ctggagagat acagctgggt cagtaggact
901 gggacaggca gctggagaat tgccatgtag atgttcatac aatcgtcaaa tcatgaaggc
961 tggaaaagcc ctccaagatc cccaagacca accccaaccc acccaccgtg cccactggcc
1021 atgtccctca gtgccacatc cccacagttc ttcatcacct ccagggacgg tgaccccccc
1081 acctccgtgg gcagctgtgc cactgcagca ccgctctttg gagaaggtaa atcttgctaa
1141 atccagcccg accctcccct ggcacaacgt aaggccatta tctctcatcc aactccagga
1201 cggagtcagt gagaatattg cctccctggc gagctcacgg ggacagcccc cccccaaagc
1261 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc gcccggggct
1321 ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg
1381 gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc
1441 tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga tttagaatga
1501 cagaatcata gaacggcctg ggttgcaaag gagcacagtg ctcatccaga tccaaccccc
1561 tgctatgtgc agggtcatca accagcagcc caggctgccc agagccacat ccagcctggc
1621 cttgaatgcc tgcagggatg gggcatccac agcctccttg ggcaacctgt tcagtgcgtc
1681 accaccctct gggggaaaaa ctgcctcctc atatccaacc caaacctccc ctgtctcagt
1741 gtaaagccat tcccccttgt cctatcaagg gggagtttgc tgtgacattg ttggtctggg
1801 gtgacacatg tttgccaatt cagtgcatca cggagaggca gatcttgggg ataaggaagt
1861 gcaggacagc atggacgtgg gacatgcagg tgttgagggc tctgggacac tctccaagtc
1921 acagcgttca gaacagcctt aaggataaga agataggata gaaggacaaa gagcaagtta
1981 aaacccagca tggagaggag cacaaaaagg ccacagacac tgctggtccc tgtgtctgag
2041 cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt ggaagagctt gcctggagag
2101 atacagctgg gtcagtagga ctgggacagg cagctggaga attgccatgt agatgttcat
2161 acaatcgtca aatcatgaag gctggaaaag ccctccaaga tccccaagac caaccccaac
2221 ccacccaccg tgcccactgg ccatgtccct cagtgccaca tccccacagt tcttcatcac
2281 ctccagggac ggtgaccccc ccacctccgt gggcagctgt gccactgcag caccgctctt
2341 tggagaaggt aaatcttgct aaatccagcc cgaccctccc ctggcacaac gtaaggccat
2401 tatctctcat ccaactccag gacggagtca gtgagaatat tcggccgctc tagaactagt
2461 ggatcccccg ggctgcagga attcgatggc cgcttgggcg ggatatctag cgccaaggtt
2521 ccgtaaagct ctctattaga ggactggcgg tgccccatca ccctgggagg cctctccaca
2581 aatatcggct acttccaatt gattggacgc gccatcttgt ctgcttatgc atattcagag
2641 gatcctgaat attcatgagc gagggccgtg cggcccctcc ctccaaccct cccccggaac
2701 aagctccgga gaacccgaca ggccccgcct tctttaccga tgcgtagaac aaaccatttt
2761 ccgggttggg gggggaaatt aatgagagac tttagctgaa aatgagcatg gacgccaaag
2821 ctgagtaaag atggcttaac tttatcctcc attctgtaat ccgtcagctt gagtgtacgg
2881 gaagtcggcg aggggggcgg caggctcaga aacattctcc tcctcctgtc gcgtcagaaa
2941 gaacacccaa ccagggagcc ggagccctgg cgtcaacatc tctggcgcgc gcgctccatg
3001 taggccggtg cgggcggccc cgtagcgcaa gggagggcgg gaaaggaagg ggcgggacac
3061 aagggcgaat ctataaaggg cgtcattcag ccagttctct cctcagaagc gccgagagcg
3121 cgaccgggac ggttggagaa gaaggtggct cccggaaggg ggagagacaa actgccgtaa
3181 cctctgccgt tcaggatccc ggccatcaag cttatcgatg ctgccgcagc aaaagcagga
3241 gcagatgccg ccgtcgcagg cgaagatgtc gcagacggag gaggcgatgc tgccggcgga
3301 ggaggcgaag taagtagagg gctgggctgg gctgtggggg gtgtggggtg cgggactggg
3361 cagtctggga gtccctctca ccacttttct tacctttcta ggatgctgcc tcgaggaact
3421 gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga
3481 tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc
3541 ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat tgtacccaat tcgttaaggc
3601 caaattggcc accatgggct ctagcctgga cgacgagcac atcctgagcg ccctgctgca
3661 gagcgacgac gaactggtgg gcgaggacag cgacagcgag gtcagcgacc acgtgtccga
3721 ggacgacgtg cagtccgaca ccgaggaagc cttcatcgac gaggtgcacg aagtgcagcc
3781 taccagcagc ggctccgaga tcctggacga gcagaacgtg atcgagcagc ctggcagctc
3841 cctggccagc aacagaatcc tgaccctgcc ccagagaacc atcagaggca agaacaagca
3901 ctgctggtcc acctccaaga gcaccaggcg gagcagagtg tccgccctga acatcgtgcg
3961 gagccagagg ggccccacca gaatgtgcag aaacatctac gaccccctgc tgtgcttcaa
4021 gctgttcttc accgacgaga tcatcagcga gatcgtgaag tggaccaacg ccgagatcag
4081 cctgaagagg cgggagagca tgaccagcgc caccttcaga gacaccaacg aggacgagat
4141 ctacgccttc ttcggcatcc tggtgatgac cgccgtgaga aaggacaacc acatgagcac
4201 cgacgacctg ttcgacagat ccctgagcat ggtgtacgtg tccgtgatga gcagagacag
4261 attcgacttc ctgatcagat gcctgagaat ggacgacaag agcatcagac ccaccctgcg
4321 ggagaacgac gtgttcaccc ccgtgcggaa gatctgggac ctgttcatcc accagtgcat
4381 ccagaactac acccctggcg cccacctgac catcgatgag cagctgctgg gcttcagagg
4441 cagatgcccc ttcagagtgt acatccccaa caagcccagc aagtacggca tcaagatcct
4501 gatgatgtgc gacagcggca ccaagtacat gatcaacggc atgccctacc tgggcagagg
4561 cacccagaca aacggcgtgc ccctgggcga gtactacgtg aaagaactga gcaagcctgt
4621 gcatggcagc tgcaggaaca tcacctgcga caactggttc accagcatcc ccctggccaa
4681 gaacctgctg caggaaccct acaagctgac catcgtgggc accgtgcgga gcaacaagcg
4741 ggagatccca gaggtgctga agaacagcag atccagacct gtgggaacaa gcatgttctg
4801 cttcgacggc cccctgaccc tggtgtccta caagcccaag cccgccaaga tggtgtacct
4861 gctgtccagc tgcgacgagg acgccagcat caacgagagc accggcaagc cccagatggt
4921 gatgtactac aaccagacca agggcggcgt ggacaccctg gaccagatgt gcagcgtgat
4981 gacctgcagc agaaagacca acagatggcc catggccctg ctgtacggca tgatcaatat
5041 cgcctgcatc aacagcttca tcatctacag ccacaacgtg tccagcaagg gcgagaaggt
5101 gcagagccgg aagaaattca tgcggaacct gtacatgagc ctgacctcca gcttcatgag
5161 aaagagactg gaagccccca ccctgaagag atacctgcgg gacaacatca gcaacatcct
5221 gcccaaggaa gtgccaggaa caagcgacga cagcaccgag gaacccgtga tgaagaagag
5281 gacctactgc acctactgtc ccagcaagat cagaagaaag gccaacgcca gctgcaagaa
5341 atgcaaaaaa gtgatctgcc gggagcacaa catcgacatg tgccagagct gtttctgagg
5401 ccgtaacggc cgccagaatt ggggatccag acatgataag atacattgat gagtttggac
5461 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
5521 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
5581 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgacct
5641 gcaggcatgc aagctcggta cccagctttt gttcccttta gtgagggtta attgcgcgct
5701 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
5761 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
5821 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
5881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
5941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
6001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
6061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
6121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
6181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
6241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
6301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
6361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
6421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
6481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
6541 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
6601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
6661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
6721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
6781 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
6841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
6901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
6961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
7021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
7081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
7141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
7201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
7261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
7321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
7381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
7441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
7501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
7561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
7621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
7681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
7741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
7801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
7861 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
7921 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
7981 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
8041 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
8101 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
8161 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
8221 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
8281 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
8341 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
8401 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
8461 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct
8521 ccaccgcggt g
//
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
| P4706/pCMV-SPORT6-PDS5A人源基因质粒 |
| P4707/pCMV-SPORT6-SLC35A5人源基因质粒 |
| P4708/pCMV-SPORT6-KCNN4人源基因质粒 |
| P4709/pCMV-SPORT6-MICA（2点突变）人源基因质粒 |
| P4710/pCMV-SPORT6-KIAA1191人源基因质粒 |
| P4711/pCMV-SPORT6-GRN人源基因质粒 |
| P4712/pCMV-SPORT6-SACM1L人源基因质粒 |
| P4713/pCMV-SPORT6-GDAP2人源基因质粒 |
| P4714/pCMV-SPORT6-SAPS3人源基因质粒 |
| P4715/pCMV-SPORT6-YWHAG人源基因质粒 |
| P4716/pCMV-SPORT6-GNPDA2人源基因质粒 |
| P4717/pCMV-SPORT6-TMEM99人源基因质粒 |
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文献和实验相关实验
Sleeping Beauty (SB) transposon-based transfection is a two-component system consisting of a transposase and a transposon containing inverted repeat/direct repeat (IR/DR) sequences that result in precise integration into a TA dinucleotide. The transposon
Super PiggyBac Transposase (PB200PA-1) 哺乳转座质粒
￥100 - 1000
