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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：
pLVX-Tet-On-Advanced
- 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：pLVX-Tet-On-Advanced慢病毒表达质粒
别称: pLVX-Tet-On-Advanced
质粒属性
质粒简介
pLVX-Tet-On-Advanced是Tet-On系统的调控载体，与之对应的应答载体是pLVX-Tight-puro.
质粒图谱
质粒序列
LOCUS Exported 11625 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 27
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11625)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..11625
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2797..3543
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/note="rtTA-Advanced"
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 3570..4143
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4157..4951
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPAT-CPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 5147..5735
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5618..5629)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5942..6575
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
polyA_signal 6688..6912
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 6958..7386
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7400..7729
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 7580..7715
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 7778..8803
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 8933..9054
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(9103..9119)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9127..9143
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(9151..9181)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9196..9217
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9505..10093)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10264..11124)
/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(11125..11229)
/gene="bla"
/note="AmpR promoter"
polyA_signal 11277..11411
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tccaccatgt ctagactgga caagagcaaa
2821 gtcataaacg gcgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg
2881 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
2941 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
3001 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt
3061 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
3121 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
3181 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
3241 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
3301 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
3361 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
3421 gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
3481 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
3541 taactagaat tccgcccccc cccccccccc ccctctccct cccccccccc taacgttact
3601 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
3661 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
3721 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
3781 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
3841 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
3901 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
3961 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
4021 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
4081 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
4141 taagcttgcc acaaccatgg ctgaacaaga tggattgcac gcaggttctc cggccgcttg
4201 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
4261 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
4321 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
4381 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
4441 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
4501 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
4561 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
4621 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
4681 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
4741 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
4801 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
4861 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
4921 cttctatcgc cttcttgacg agttcttctg agcgggatcg gctagagtcg acccggacgg
4981 gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc
5041 cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag
5101 ctgctcacca cccactcgag tctagagggc ccgacgcgtc tggaacaatc aacctctgga
5161 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
5221 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
5281 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
5341 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
5401 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
5461 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
5521 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
5581 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
5641 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
5701 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
5761 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
5821 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
5881 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
5941 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
6001 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
6061 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
6121 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
6181 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
6241 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
6301 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
6361 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
6421 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
6481 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
6541 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
6601 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
6661 gcgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
6721 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
6781 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
6841 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
6901 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
6961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
7021 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
7081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
7141 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
7201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
7261 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
7321 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
7381 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
7441 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
7501 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
7561 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
7621 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
7681 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
7741 agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg aactcaccgc
7801 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
7861 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
7921 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
7981 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
8041 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
8101 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
8161 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
8221 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
8281 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
8341 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
8401 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
8461 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
8521 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
8581 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
8641 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
8701 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
8761 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt
8821 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
8881 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
8941 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
9001 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
9061 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
9121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
9181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
9241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
9301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
9361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
9421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
9481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
9541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
9601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
9661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
9721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
9781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
9841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
9901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
9961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
10021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
10081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
10141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
10201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
10261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
10321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
10381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
10441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
10501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
10561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
10621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
10681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
10741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
10801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
10861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
10921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
10981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
11041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
11101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
11161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
11221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact
11281 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
11341 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
11401 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
11461 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
11521 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt
11581 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
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：pLVX-Tet-On-Advanced慢病毒表达质粒
别称: pLVX-Tet-On-Advanced
| 启动子: | CMV |
|---|---|
| 复制子: | pUC |
| 终止子: | SV40 poly(A) signal,bGH poly(A) signal |
| 质粒分类: | 病毒系列，慢病毒克隆载体 |
| 质粒大小: | 11,625bp |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37度 |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 四环素诱导 |
| 5'测序引物: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
| 3'测序引物: | 根据序列设计引物 |
质粒属性
| 质粒宿主： | 哺乳细胞,慢病毒 |
|---|---|
| 质粒用途： | 诱导,辅助包装 |
| 片段类型： | ORF |
| 片段物种： | |
| 原核抗性： | Amp |
| 筛选标记： | Hyg |
| 荧光标记： |
质粒简介
pLVX-Tet-On-Advanced是Tet-On系统的调控载体，与之对应的应答载体是pLVX-Tight-puro.
质粒图谱
质粒序列
LOCUS Exported 11625 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 27
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11625)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..11625
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2797..3543
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/note="rtTA-Advanced"
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 3570..4143
/note="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4157..4951
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPAT-CPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 5147..5735
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5618..5629)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 5942..6575
/note="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
polyA_signal 6688..6912
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 6958..7386
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7400..7729
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 7580..7715
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 7778..8803
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMD-AIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 8933..9054
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(9103..9119)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9127..9143
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(9151..9181)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9196..9217
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9505..10093)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10264..11124)
/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(11125..11229)
/gene="bla"
/note="AmpR promoter"
polyA_signal 11277..11411
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tccaccatgt ctagactgga caagagcaaa
2821 gtcataaacg gcgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg
2881 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
2941 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
3001 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt
3061 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
3121 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
3181 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
3241 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
3301 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
3361 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
3421 gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
3481 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
3541 taactagaat tccgcccccc cccccccccc ccctctccct cccccccccc taacgttact
3601 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
3661 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
3721 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
3781 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
3841 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
3901 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
3961 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
4021 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
4081 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
4141 taagcttgcc acaaccatgg ctgaacaaga tggattgcac gcaggttctc cggccgcttg
4201 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
4261 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
4321 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
4381 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
4441 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
4501 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
4561 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
4621 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
4681 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
4741 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
4801 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
4861 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
4921 cttctatcgc cttcttgacg agttcttctg agcgggatcg gctagagtcg acccggacgg
4981 gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc
5041 cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag
5101 ctgctcacca cccactcgag tctagagggc ccgacgcgtc tggaacaatc aacctctgga
5161 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
5221 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
5281 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
5341 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
5401 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
5461 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
5521 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
5581 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
5641 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
5701 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
5761 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
5821 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
5881 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
5941 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
6001 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
6061 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
6121 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
6181 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
6241 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
6301 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
6361 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
6421 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
6481 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
6541 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
6601 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
6661 gcgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
6721 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
6781 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
6841 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
6901 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
6961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
7021 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
7081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
7141 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
7201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
7261 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
7321 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
7381 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
7441 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
7501 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
7561 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
7621 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
7681 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
7741 agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg aactcaccgc
7801 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
7861 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
7921 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
7981 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
8041 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
8101 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
8161 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
8221 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
8281 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
8341 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
8401 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
8461 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
8521 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
8581 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
8641 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
8701 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
8761 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt
8821 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
8881 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
8941 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
9001 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
9061 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
9121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
9181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
9241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
9301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
9361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
9421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
9481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
9541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
9601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
9661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
9721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
9781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
9841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
9901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
9961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
10021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
10081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
10141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
10201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
10261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
10321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
10381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
10441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
10501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
10561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
10621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
10681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
10741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
10801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
10861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
10921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
10981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
11041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
11101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
11161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
11221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact
11281 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
11341 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
11401 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
11461 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
11521 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt
11581 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
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
| P3634/pCMV-SPORT6-STX8（2同义突变）人源基因质粒 |
| P3635/pCMV-SPORT6-APOC1人源基因质粒 |
| P3636/pCMV-SPORT6-GNA15人源基因质粒 |
| P3637/pCMV-SPORT6-RPL18人源基因质粒 |
| P3638/pCMV-SPORT6-NEIL1（1点突变）人源基因质粒 |
| P3639/pCMV-SPORT6-SLC47A1人源基因质粒 |
| P3640/pCMV-SPORT6-PRKCSH人源基因质粒 |
| P3641/pCMV-SPORT6-TM7SF2(1同义突变）人源基因质粒 |
| P3642/pCMV-SPORT6-SPAG16人源基因质粒 |
| P3643/pCMV-SPORT6-LMO4人源基因质粒 |
| P3644/pCMV-SPORT6-HVCN1人源基因质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验相关实验
Amp pLVX-DsRed-Monomer-N1 病毒质粒 Amp pLVX-Tight-Puro 病毒质粒 pLVX-Tet-On-Advanced 病毒
pLVX-Tet-On-Advanced慢病毒表达质粒
￥100 - 1000
