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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：
Plasmid#12260;gag-pol
- 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：psPAX2慢病毒包装质粒
别称: Plasmid#12260;gag-pol
质粒属性
质粒简介
2nd generation packaging vector. Known to work with many of the Aebischer and Trono lab lentiviral vectors, as well as the pLKO system. Produces higher titer than pCMV-dR8.2 dvpr.
质粒图谱
质粒序列
LOCUS Exported 10703 bp ds-DNA circular SYN 02-MAY-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10703
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/note="chicken beta-actin promoter"
intron 662..1670
/note="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 1780..3282
/codon_start=1
/gene="gag"
/product="gag protein from human immunodeficiency virus 1"
/note="HIV-1 gag"
/translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 3075..6086
/codon_start=1
/gene="pol"
/product="pol protein from human immunodeficiency virus 1"
/note="HIV-1 pol"
/translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
GIGGFIKVGQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
STKWRKLVDFRELNKRTQDFWEVQLGIP-HPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
KGIGGNEQVDGLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
MAGDDCVASRQDED"
misc_feature 6660..6893
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
polyA_site 7792..7847
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
promoter 8379..8574
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 8425..8560
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 8580..8714
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(8953..9541)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9712..10572)
/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(10573..10677)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgggccg gccgcgttga
1741 cgcgcacggc aagaggcgag gggcggcgac tggtgagaga tgggtgcgag agcgtcagta
1801 ttaagcgggg gagaattaga tcgatgggaa aaaattcggt taaggccagg gggaaagaaa
1861 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
1921 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
1981 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
2041 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
2101 caaaacaaaa gtaagaaaaa agcacagcaa gcagcagctg acacaggaca cagcaatcag
2161 gtcagccaaa attaccctat agtgcagaac atccaggggc aaatggtaca tcaggccata
2221 tcacctagaa ctttaaatgc atgggtaaaa gtagtagaag agaaggcttt cagcccagaa
2281 gtgataccca tgttttcagc attatcagaa ggagccaccc cacaagattt aaacaccatg
2341 ctaaacacag tggggggaca tcaagcagcc atgcaaatgt taaaagagac catcaatgag
2401 gaagctgcag aatgggatag agtgcatcca gtgcatgcag ggcctattgc accaggccag
2461 atgagagaac caaggggaag tgacatagca ggaactacta gtacccttca ggaacaaata
2521 ggatggatga cacataatcc acctatccca gtaggagaaa tctataaaag atggataatc
2581 ctgggattaa ataaaatagt aagaatgtat agccctacca gcattctgga cataagacaa
2641 ggaccaaagg aaccctttag agactatgta gaccgattct ataaaactct aagagccgag
2701 caagcttcac aagaggtaaa aaattggatg acagaaacct tgttggtcca aaatgcgaac
2761 ccagattgta agactatttt aaaagcattg ggaccaggag cgacactaga agaaatgatg
2821 acagcatgtc agggagtggg gggacccggc cataaagcaa gagttttggc tgaagcaatg
2881 agccaagtaa caaatccagc taccataatg atacagaaag gcaattttag gaaccaaaga
2941 aagactgtta agtgtttcaa ttgtggcaaa gaagggcaca tagccaaaaa ttgcagggcc
3001 cctaggaaaa agggctgttg gaaatgtgga aaggaaggac accaaatgaa agattgtact
3061 gagagacagg ctaatttttt agggaagatc tggccttccc acaagggaag gccagggaat
3121 tttcttcaga gcagaccaga gccaacagcc ccaccagaag agagcttcag gtttggggaa
3181 gagacaacaa ctccctctca gaagcaggag ccgatagaca aggaactgta tcctttagct
3241 tccctcagat cactctttgg cagcgacccc tcgtcacaat aaagataggg gggcaattaa
3301 aggaagctct attagataca ggagcagatg atacagtatt agaagaaatg aatttgccag
3361 gaagatggaa accaaaaatg atagggggaa ttggaggttt tatcaaagta ggacagtatg
3421 atcagatact catagaaatc tgcggacata aagctatagg tacagtatta gtaggaccta
3481 cacctgtcaa cataattgga agaaatctgt tgactcagat tggctgcact ttaaattttc
3541 ccattagtcc tattgagact gtaccagtaa aattaaagcc aggaatggat ggcccaaaag
3601 ttaaacaatg gccattgaca gaagaaaaaa taaaagcatt agtagaaatt tgtacagaaa
3661 tggaaaagga aggaaaaatt tcaaaaattg ggcctgaaaa tccatacaat actccagtat
3721 ttgccataaa gaaaaaagac agtactaaat ggagaaaatt agtagatttc agagaactta
3781 ataagagaac tcaagatttc tgggaagttc aattaggaat accacatcct gcagggttaa
3841 aacagaaaaa atcagtaaca gtactggatg tgggcgatgc atatttttca gttcccttag
3901 ataaagactt caggaagtat actgcattta ccatacctag tataaacaat gagacaccag
3961 ggattagata tcagtacaat gtgcttccac agggatggaa aggatcacca gcaatattcc
4021 agtgtagcat gacaaaaatc ttagagcctt ttagaaaaca aaatccagac atagtcatct
4081 atcaatacat ggatgatttg tatgtaggat ctgacttaga aatagggcag catagaacaa
4141 aaatagagga actgagacaa catctgttga ggtggggatt taccacacca gacaaaaaac
4201 atcagaaaga acctccattc ctttggatgg gttatgaact ccatcctgat aaatggacag
4261 tacagcctat agtgctgcca gaaaaggaca gctggactgt caatgacata cagaaattag
4321 tgggaaaatt gaattgggca agtcagattt atgcagggat taaagtaagg caattatgta
4381 aacttcttag gggaaccaaa gcactaacag aagtagtacc actaacagaa gaagcagagc
4441 tagaactggc agaaaacagg gagattctaa aagaaccggt acatggagtg tattatgacc
4501 catcaaaaga cttaatagca gaaatacaga agcaggggca aggccaatgg acatatcaaa
4561 tttatcaaga gccatttaaa aatctgaaaa caggaaaata tgcaagaatg aagggtgccc
4621 acactaatga tgtgaaacaa ttaacagagg cagtacaaaa aatagccaca gaaagcatag
4681 taatatgggg aaagactcct aaatttaaat tacccataca aaaggaaaca tgggaagcat
4741 ggtggacaga gtattggcaa gccacctgga ttcctgagtg ggagtttgtc aatacccctc
4801 ccttagtgaa gttatggtac cagttagaga aagaacccat aataggagca gaaactttct
4861 atgtagatgg ggcagccaat agggaaacta aattaggaaa agcaggatat gtaactgaca
4921 gaggaagaca aaaagttgtc cccctaacgg acacaacaaa tcagaagact gagttacaag
4981 caattcatct agctttgcag gattcgggat tagaagtaaa catagtgaca gactcacaat
5041 atgcattggg aatcattcaa gcacaaccag ataagagtga atcagagtta gtcagtcaaa
5101 taatagagca gttaataaaa aaggaaaaag tctacctggc atgggtacca gcacacaaag
5161 gaattggagg aaatgaacaa gtagatgggt tggtcagtgc tggaatcagg aaagtactat
5221 ttttagatgg aatagataag gcccaagaag aacatgagaa atatcacagt aattggagag
5281 caatggctag tgattttaac ctaccacctg tagtagcaaa agaaatagta gccagctgtg
5341 ataaatgtca gctaaaaggg gaagccatgc atggacaagt agactgtagc ccaggaatat
5401 ggcagctaga ttgtacacat ttagaaggaa aagttatctt ggtagcagtt catgtagcca
5461 gtggatatat agaagcagaa gtaattccag cagagacagg gcaagaaaca gcatacttcc
5521 tcttaaaatt agcaggaaga tggccagtaa aaacagtaca tacagacaat ggcagcaatt
5581 tcaccagtac tacagttaag gccgcctgtt ggtgggcggg gatcaagcag gaatttggca
5641 ttccctacaa tccccaaagt caaggagtaa tagaatctat gaataaagaa ttaaagaaaa
5701 ttataggaca ggtaagagat caggctgaac atcttaagac agcagtacaa atggcagtat
5761 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
5821 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
5881 aaaattttcg ggtttattac agggacagca gagatccagt ttggaaagga ccagcaaagc
5941 tcctctggaa aggtgaaggg gcagtagtaa tacaagataa tagtgacata aaagtagtgc
6001 caagaagaaa agcaaagatc atcagggatt atggaaaaca gatggcaggt gatgattgtg
6061 tggcaagtag acaggatgag gattaacaca tggaattctg caacaactgc tgtttatcca
6121 tttcagaatt gggtgtcgac atagcagaat aggcgttact cgacagagga gagcaagaaa
6181 tggagccagt agatcctaga ctagagccct ggaagcatcc aggaagtcag cctaaaactg
6241 cttgtaccaa ttgctattgt aaaaagtgtt gctttcattg ccaagtttgt ttcatgacaa
6301 aagccttagg catctcctat ggcaggaaga agcggagaca gcgacgaaga gctcatcaga
6361 acagtcagac tcatcaagct tctctatcaa agcagtaagt agtacatgta atgcaaccta
6421 taatagtagc aatagtagca ttagtagtag caataataat agcaatagtt gtgtggtcca
6481 tagtaatcat agaatatagg aaaatggccg ctgatcttca gacctggagg aggagatatg
6541 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
6601 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
6661 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg
6721 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
6781 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
6841 ctccaagcaa gaatcctagc tgtggaaaga tacctaaagg atcaacagct cctagggatt
6901 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
6961 aataaatctc tggaacagat ctggaatcac acgacctgga tggagtggga cagagaaatt
7021 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
7081 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
7141 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
7201 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
7261 tcgtttcaga cccacctccc aatcccgagg ggacccgaca ggcccgaagg aatagaagaa
7321 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc cttggcactt
7381 atctgggacg atctgcggag cctgtgcctc ttcagctacc accgcttgag agacttactc
7441 ttgattgtaa cgaggattgt ggaacttctg ggacgcaggg ggtgggaagc cctcaaatat
7501 tggtggaatc tcctacaata ttggagtcag gagctaaaga atagtgctgt tagcttgctc
7561 aatgccacag ccatagcagt agctgagggg acagataggg ttatagaagt agtacaagga
7621 gcttgtagag ctattcgcca catacctaga agaataagac agggcttgga aaggattttg
7681 ctataagctc gaaacaaccg gtacctctag aactatagct agcagatctt tttccctctg
7741 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
7801 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
7861 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
7921 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
7981 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
8041 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
8101 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
8161 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
8221 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
8281 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
8341 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
8401 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
8461 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
8521 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
8581 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
8641 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
8701 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
8761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
8821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
8881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
8941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
9001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
9061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
9121 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
9181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
9241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
9301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
9361 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
9421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
9481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
9541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
9601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
9661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
9721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
9781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
9841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
9901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
9961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
10021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
10081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
10141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
10201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
10261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
10321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
10381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
10441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
10501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
10561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
10621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
10681 atttccccga aaagtgccac ctg
//
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：psPAX2慢病毒包装质粒
别称: Plasmid#12260;gag-pol
| 启动子: | CAG |
|---|---|
| 复制子: | pUC |
| 终止子: | β-globin poly(A) signal,SV40 poly(A) signal |
| 质粒分类: | 病毒系列，慢病毒包装载体 |
| 原核抗性: | Amp |
| 克隆菌株: | DH5a |
| 培养条件: | 37度 |
| 表达宿主: | 哺乳细胞293T |
| 诱导方式: | 无须诱导，瞬时表达 |
| 5'测序引物: | pcaggs-F |
| 3'测序引物: | pcaggs-R |
质粒属性
| 载体宿主： | 哺乳细胞,慢病毒 |
|---|---|
| 载体用途： | 包装辅助 |
| 基因种属： | 病毒 |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： |
质粒简介
2nd generation packaging vector. Known to work with many of the Aebischer and Trono lab lentiviral vectors, as well as the pLKO system. Produces higher titer than pCMV-dR8.2 dvpr.
质粒图谱
质粒序列
LOCUS Exported 10703 bp ds-DNA circular SYN 02-MAY-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10703
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/note="chicken beta-actin promoter"
intron 662..1670
/note="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 1780..3282
/codon_start=1
/gene="gag"
/product="gag protein from human immunodeficiency virus 1"
/note="HIV-1 gag"
/translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 3075..6086
/codon_start=1
/gene="pol"
/product="pol protein from human immunodeficiency virus 1"
/note="HIV-1 pol"
/translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
GIGGFIKVGQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
STKWRKLVDFRELNKRTQDFWEVQLGIP-HPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
KGIGGNEQVDGLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
MAGDDCVASRQDED"
misc_feature 6660..6893
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
polyA_site 7792..7847
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
promoter 8379..8574
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 8425..8560
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 8580..8714
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(8953..9541)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9712..10572)
/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(10573..10677)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgggccg gccgcgttga
1741 cgcgcacggc aagaggcgag gggcggcgac tggtgagaga tgggtgcgag agcgtcagta
1801 ttaagcgggg gagaattaga tcgatgggaa aaaattcggt taaggccagg gggaaagaaa
1861 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
1921 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
1981 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
2041 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
2101 caaaacaaaa gtaagaaaaa agcacagcaa gcagcagctg acacaggaca cagcaatcag
2161 gtcagccaaa attaccctat agtgcagaac atccaggggc aaatggtaca tcaggccata
2221 tcacctagaa ctttaaatgc atgggtaaaa gtagtagaag agaaggcttt cagcccagaa
2281 gtgataccca tgttttcagc attatcagaa ggagccaccc cacaagattt aaacaccatg
2341 ctaaacacag tggggggaca tcaagcagcc atgcaaatgt taaaagagac catcaatgag
2401 gaagctgcag aatgggatag agtgcatcca gtgcatgcag ggcctattgc accaggccag
2461 atgagagaac caaggggaag tgacatagca ggaactacta gtacccttca ggaacaaata
2521 ggatggatga cacataatcc acctatccca gtaggagaaa tctataaaag atggataatc
2581 ctgggattaa ataaaatagt aagaatgtat agccctacca gcattctgga cataagacaa
2641 ggaccaaagg aaccctttag agactatgta gaccgattct ataaaactct aagagccgag
2701 caagcttcac aagaggtaaa aaattggatg acagaaacct tgttggtcca aaatgcgaac
2761 ccagattgta agactatttt aaaagcattg ggaccaggag cgacactaga agaaatgatg
2821 acagcatgtc agggagtggg gggacccggc cataaagcaa gagttttggc tgaagcaatg
2881 agccaagtaa caaatccagc taccataatg atacagaaag gcaattttag gaaccaaaga
2941 aagactgtta agtgtttcaa ttgtggcaaa gaagggcaca tagccaaaaa ttgcagggcc
3001 cctaggaaaa agggctgttg gaaatgtgga aaggaaggac accaaatgaa agattgtact
3061 gagagacagg ctaatttttt agggaagatc tggccttccc acaagggaag gccagggaat
3121 tttcttcaga gcagaccaga gccaacagcc ccaccagaag agagcttcag gtttggggaa
3181 gagacaacaa ctccctctca gaagcaggag ccgatagaca aggaactgta tcctttagct
3241 tccctcagat cactctttgg cagcgacccc tcgtcacaat aaagataggg gggcaattaa
3301 aggaagctct attagataca ggagcagatg atacagtatt agaagaaatg aatttgccag
3361 gaagatggaa accaaaaatg atagggggaa ttggaggttt tatcaaagta ggacagtatg
3421 atcagatact catagaaatc tgcggacata aagctatagg tacagtatta gtaggaccta
3481 cacctgtcaa cataattgga agaaatctgt tgactcagat tggctgcact ttaaattttc
3541 ccattagtcc tattgagact gtaccagtaa aattaaagcc aggaatggat ggcccaaaag
3601 ttaaacaatg gccattgaca gaagaaaaaa taaaagcatt agtagaaatt tgtacagaaa
3661 tggaaaagga aggaaaaatt tcaaaaattg ggcctgaaaa tccatacaat actccagtat
3721 ttgccataaa gaaaaaagac agtactaaat ggagaaaatt agtagatttc agagaactta
3781 ataagagaac tcaagatttc tgggaagttc aattaggaat accacatcct gcagggttaa
3841 aacagaaaaa atcagtaaca gtactggatg tgggcgatgc atatttttca gttcccttag
3901 ataaagactt caggaagtat actgcattta ccatacctag tataaacaat gagacaccag
3961 ggattagata tcagtacaat gtgcttccac agggatggaa aggatcacca gcaatattcc
4021 agtgtagcat gacaaaaatc ttagagcctt ttagaaaaca aaatccagac atagtcatct
4081 atcaatacat ggatgatttg tatgtaggat ctgacttaga aatagggcag catagaacaa
4141 aaatagagga actgagacaa catctgttga ggtggggatt taccacacca gacaaaaaac
4201 atcagaaaga acctccattc ctttggatgg gttatgaact ccatcctgat aaatggacag
4261 tacagcctat agtgctgcca gaaaaggaca gctggactgt caatgacata cagaaattag
4321 tgggaaaatt gaattgggca agtcagattt atgcagggat taaagtaagg caattatgta
4381 aacttcttag gggaaccaaa gcactaacag aagtagtacc actaacagaa gaagcagagc
4441 tagaactggc agaaaacagg gagattctaa aagaaccggt acatggagtg tattatgacc
4501 catcaaaaga cttaatagca gaaatacaga agcaggggca aggccaatgg acatatcaaa
4561 tttatcaaga gccatttaaa aatctgaaaa caggaaaata tgcaagaatg aagggtgccc
4621 acactaatga tgtgaaacaa ttaacagagg cagtacaaaa aatagccaca gaaagcatag
4681 taatatgggg aaagactcct aaatttaaat tacccataca aaaggaaaca tgggaagcat
4741 ggtggacaga gtattggcaa gccacctgga ttcctgagtg ggagtttgtc aatacccctc
4801 ccttagtgaa gttatggtac cagttagaga aagaacccat aataggagca gaaactttct
4861 atgtagatgg ggcagccaat agggaaacta aattaggaaa agcaggatat gtaactgaca
4921 gaggaagaca aaaagttgtc cccctaacgg acacaacaaa tcagaagact gagttacaag
4981 caattcatct agctttgcag gattcgggat tagaagtaaa catagtgaca gactcacaat
5041 atgcattggg aatcattcaa gcacaaccag ataagagtga atcagagtta gtcagtcaaa
5101 taatagagca gttaataaaa aaggaaaaag tctacctggc atgggtacca gcacacaaag
5161 gaattggagg aaatgaacaa gtagatgggt tggtcagtgc tggaatcagg aaagtactat
5221 ttttagatgg aatagataag gcccaagaag aacatgagaa atatcacagt aattggagag
5281 caatggctag tgattttaac ctaccacctg tagtagcaaa agaaatagta gccagctgtg
5341 ataaatgtca gctaaaaggg gaagccatgc atggacaagt agactgtagc ccaggaatat
5401 ggcagctaga ttgtacacat ttagaaggaa aagttatctt ggtagcagtt catgtagcca
5461 gtggatatat agaagcagaa gtaattccag cagagacagg gcaagaaaca gcatacttcc
5521 tcttaaaatt agcaggaaga tggccagtaa aaacagtaca tacagacaat ggcagcaatt
5581 tcaccagtac tacagttaag gccgcctgtt ggtgggcggg gatcaagcag gaatttggca
5641 ttccctacaa tccccaaagt caaggagtaa tagaatctat gaataaagaa ttaaagaaaa
5701 ttataggaca ggtaagagat caggctgaac atcttaagac agcagtacaa atggcagtat
5761 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
5821 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
5881 aaaattttcg ggtttattac agggacagca gagatccagt ttggaaagga ccagcaaagc
5941 tcctctggaa aggtgaaggg gcagtagtaa tacaagataa tagtgacata aaagtagtgc
6001 caagaagaaa agcaaagatc atcagggatt atggaaaaca gatggcaggt gatgattgtg
6061 tggcaagtag acaggatgag gattaacaca tggaattctg caacaactgc tgtttatcca
6121 tttcagaatt gggtgtcgac atagcagaat aggcgttact cgacagagga gagcaagaaa
6181 tggagccagt agatcctaga ctagagccct ggaagcatcc aggaagtcag cctaaaactg
6241 cttgtaccaa ttgctattgt aaaaagtgtt gctttcattg ccaagtttgt ttcatgacaa
6301 aagccttagg catctcctat ggcaggaaga agcggagaca gcgacgaaga gctcatcaga
6361 acagtcagac tcatcaagct tctctatcaa agcagtaagt agtacatgta atgcaaccta
6421 taatagtagc aatagtagca ttagtagtag caataataat agcaatagtt gtgtggtcca
6481 tagtaatcat agaatatagg aaaatggccg ctgatcttca gacctggagg aggagatatg
6541 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
6601 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
6661 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg
6721 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
6781 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
6841 ctccaagcaa gaatcctagc tgtggaaaga tacctaaagg atcaacagct cctagggatt
6901 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
6961 aataaatctc tggaacagat ctggaatcac acgacctgga tggagtggga cagagaaatt
7021 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
7081 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
7141 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
7201 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
7261 tcgtttcaga cccacctccc aatcccgagg ggacccgaca ggcccgaagg aatagaagaa
7321 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc cttggcactt
7381 atctgggacg atctgcggag cctgtgcctc ttcagctacc accgcttgag agacttactc
7441 ttgattgtaa cgaggattgt ggaacttctg ggacgcaggg ggtgggaagc cctcaaatat
7501 tggtggaatc tcctacaata ttggagtcag gagctaaaga atagtgctgt tagcttgctc
7561 aatgccacag ccatagcagt agctgagggg acagataggg ttatagaagt agtacaagga
7621 gcttgtagag ctattcgcca catacctaga agaataagac agggcttgga aaggattttg
7681 ctataagctc gaaacaaccg gtacctctag aactatagct agcagatctt tttccctctg
7741 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
7801 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
7861 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
7921 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
7981 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
8041 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
8101 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
8161 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
8221 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
8281 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
8341 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
8401 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
8461 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
8521 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
8581 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
8641 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
8701 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
8761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
8821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
8881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
8941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
9001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
9061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
9121 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
9181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
9241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
9301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
9361 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
9421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
9481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
9541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
9601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
9661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
9721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
9781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
9841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
9901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
9961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
10021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
10081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
10141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
10201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
10261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
10321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
10381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
10441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
10501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
10561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
10621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
10681 atttccccga aaagtgccac ctg
//
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
| P0445/pN3哺乳表达质粒 |
| P1019/pCR3.1哺乳表达质粒 |
| P1027/pBK-CMV哺乳表达质粒 |
| P1101/pJG-ALPHA-MHC哺乳表达质粒 |
| P1149/pRC-CMV哺乳表达质粒 |
| P1750/pRK5-HA哺乳表达质粒 |
| P1300/pRK5-2×HA哺乳表达质粒 |
| P0133/pEGFP-N1哺乳荧光质粒 |
| P0153/pEGFP-N2哺乳荧光质粒 |
| P0452/pEGFP-N3哺乳荧光质粒 |
| P0134/pEGFP-C1哺乳荧光质粒 |
| P0135/pEGFP-C2哺乳荧光质粒 |
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psPAX2慢病毒包装质粒
￥100 - 1000
