• 规格：20μl质粒
• 启动子：CMV
• 复制子：pUC
• 终止子：WPRE
• 原核抗性：氨苄青霉素Amp
• 筛选标记：嘌呤霉素Puro
• 克隆菌株：大肠杆菌Stbl3
• 质粒简介：                   pLVML-3×FLAG-N6AMT1-IRES-Puro(NM_013240.5)是一个人源基因慢病毒哺乳细胞过表达质粒。
• 质粒图谱1： png/1-1G2131F420925.png

质粒序列： 点击查看序列

LOCUS       Exported                8840 bp ds-DNA     circular SYN 13-DEC-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..8840

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     LTR             1..634

                     /label=3' LTR

                     /note="3' long terminal repeat (LTR) from HIV-1"

     misc_feature    681..806

                     /label=HIV-1 Psi

                     /note="packaging signal of human immunodeficiency virus 

                     type 1"

     misc_feature    1303..1536

                     /label=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

                     /label=cPPT/CTS

                     /note="central polypurine tract and central termination 

                     sequence of HIV-1 (lacking the first T)"

     enhancer        2201..2504

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        2505..2708

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     regulatory      2809..2814

                     /regulatory_class="other"

                     /note="vertebrate consensus sequence for strong initiation 

                     of translation (Kozak, 1987)"

     CDS             2818..2883

                     /codon_start=1

                     /product="three tandem FLAG(R) epitope tags, followed by an

                     enterokinase cleavage site"

                     /label=3xFLAG

                     /translation="DYKDHDGDYKDHDIDYKDDDDK"

     CDS             2890..3534

                     /codon_start=1

                     /product="pLVML-3×FLAG-N6AMT1-IRES-Puro(NM_013240.5)"

                     /label=N6AMT1

                     /translation="MAGENFATPFHGHVGRGAFSDVYEPAEDTFLLLDALEAAAAELAG

                     VEICLEVGSGSGVVSAFLASMIGPQALYMCTDINPEAAACTLETARCNKVHIQPVITDL

                     VKGLLPRLTEKVDLLVFNPPYVVTPPQEVGSHGIEAAWAGGRNGREVMDRFFPLVPDLL

                     SPRGLFYLVTIKENNPEEILKIMKTKGLQGTTALSRQAGQETLSVLKFTKS"

     misc_feature    3543..4116

                     /label=IRES

                     /note="internal ribosome entry site (IRES) of the 

                     encephalomyocarditis virus (EMCV)"

     CDS             4149..4748

                     /codon_start=1

                     /gene="pac from Streptomyces alboniger"

                     /product="puromycin N-acetyltransferase"

                     /label=PuroR

                     /note="confers resistance to puromycin"

                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER

                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA

                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS

                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"

     misc_feature    4762..5350

                     /label=WPRE

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     LTR             5557..6190

                     /label=3' LTR

                     /note="3' long terminal repeat (LTR) from HIV-1"

     rep_origin      complement(6720..7308)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(7479..8339)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(8340..8444)

                     /gene="bla"

                     /label=AmpR promoter

     polyA_signal    8492..8626

                     /label=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 tctatttccg gtgaattcgc caccatggac

     2821 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac

     2881 aagctcgaga tggcagggga gaacttcgct acgccgttcc acgggcacgt gggccgcggc

     2941 gccttcagcg acgtgtacga gcccgcggag gacacgtttc tgcttttgga cgcgctggag

     3001 gcagcggctg ccgaactggc aggagtggaa atatgcctgg aagtagggtc agggtctggt

     3061 gtagtatctg cattcctagc ctctatgata ggccctcagg ctttgtacat gtgcactgat

     3121 atcaaccctg aggcagcagc ttgtacccta gagacagcac gctgtaacaa agttcacatt

     3181 caaccagtta ttacagattt ggtcaaaggc ttgctaccaa gattgaccga aaaagttgat

     3241 cttctggtgt ttaatccccc ctatgtagtg actccacctc aagaggtagg aagtcacgga

     3301 atagaggcag cttgggctgg tggcagaaat ggtcgggaag tcatggacag gttttttccc

     3361 ctggttccag atctcctttc accaagagga ttattctatt tagttaccat taaagaaaac

     3421 aacccagaag aaattttgaa aataatgaag acaaaaggtc tgcaaggaac cactgcactt

     3481 tccagacaag caggccaaga aactctttca gtcctcaagt tcaccaagtc ttagggatcc

     3541 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg