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載體描述

    pFUSE-Fc plasmids yield high levels of Fc-fusion proteins. The level of expression is usually in the μg/mL range. They can be transfected in a variety of mammalian cells, including myeloma cell lines, CHO cells, monkey COS cells and human embryonic kidney (HEK)293 cells, cells that are commonly used in protein purification systems.

pFUSE-Fc2 (IL2ss) plasmids allow the secretion of Fc-Fusion proteins.They contain the IL2 signal sequence (IL2ss) for the generation of Fc-Fusion proteins derived from proteins that are not naturally secreted. As Fc-Fusion proteins are secreted, they can be easily detected in the supernatant of pFUSE-Fc-transfected cells by SDS-PAGE. Furthermore, functional domains can be identified by immunoblotting and ligand blotting.Fc-Fusion proteins can be easily purified by single-step protein A or protein G affinity chromatography.

 載體序列

LOCUS       Exported                4194 bp ds-DNA     circular SYN 19-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4194)

  AUTHORS   BT-lab

  TITLE     Direct Submission

  JOURNAL   Exported Tuesday, Mar 19, 2019 from SnapGene Viewer 4.3.4

            http://bt-lab.cn/

FEATURES             Location/Qualifiers

     source          1..4194

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        31..242

                     /label=EF-1-alpha core promoter

                     /note="core promoter for human elongation factor

                     EF-1-alpha"

     LTR             255..523

                     /label=5' LTR (truncated)

                     /note="truncated 5' long terminal repeat (LTR) from human

                     T-cell leukemia virus (HTLV) type 1"

     sig_peptide     573..632

                     /product="human interleukin-2 signal sequence"

                     /label=IL-2 signal sequence

     misc_feature    633..653

                     /label=MCS

                     /note="Multiple cloning site"

     misc_feature    640..645

                     /label=NcoI

                     /note="NcoI restriction site: upstream insertion site for

                     scFv"

     CDS             654..1334

                     /codon_start=1

                     /product="Fc-adiponectin"

                     /product="synthetic secretion-engineered adiponectin,  

                     Synthetic construct Fc-adiponectin gene"

                     /label=Fc-adiponectin

                     /translation="DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD

                     VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS

                     NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWES

                     NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS

                     LSPGK"

     polyA_signal    1359..1480

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     polyA_signal    1584..1978

                     /label=beta-globin poly(A)

                     /note="human beta-globin polyadenylation signal"

     CDS             complement(1992..2366)

                     /codon_start=1

                     /gene="Sh ble from Streptoalloteichus hindustanus"

                     /product="antibiotic-binding protein"

                     /label=BleoR

                     /note="confers resistance to bleomycin, phleomycin, and

                     Zeocin(TM)"

                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD

                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR

                     EFALRDPAGNCVHFVAEEQD"

     promoter        complement(2416..2471)

                     /label=EM2KC promoter

                     /note="EM2KC bacterial promoter"

     promoter        complement(2589..2851)

                     /label=hFerL promoter

                     /note="human ferritin light chain mammalian cell promoter"

     enhancer        2959..3262

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     rep_origin      complement(3342..3930)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     misc_feature    4030..4037

                     /label=NotI

                     /note="NotI restriction site; downstream insertion site  

                     for scFv"

     polyA_signal    4038..4086

                     /note="synthetic polyadenylation signal"

     misc_feature    4097..4188

                     /label=pause site

                     /note="RNA polymerase II transcriptional pause signal from

                     the human alpha-2 globin gene"

ORIGIN

        1 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg

       61 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa

      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt

      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac

      241 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc

      301 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg

      361 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc

      421 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac

      481 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc

      541 ctacctgaga tcaccggcga aggagggcca ccatgtacag gatgcaactc ctgtcttgca

      601 ttgcactaag tcttgcactt gtcacgaatt cgatatcggc catggttaga tctgacaaaa

      661 ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct

      721 tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg

      781 tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg

      841 aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg

      901 tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg

      961 tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc

     1021 cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg

     1081 tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga

     1141 gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct

     1201 ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct

     1261 tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag agcctctccc

     1321 tgtctccggg taaatgagtg ctagctggcc agacatgata agatacattg atgagtttgg

     1381 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat

     1441 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca

     1501 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta

     1561 caaatgtggt atggaattaa ttctaaaata cagcatagca aaactttaac ctccaaatca

     1621 agcctctact tgaatccttt tctgagggat gaataaggca taggcatcag gggctgttgc

     1681 caatgtgcat tagctgtttg cagcctcacc ttctttcatg gagtttaaga tatagtgtat

     1741 tttcccaagg tttgaactag ctcttcattt ctttatgttt taaatgcact gacctcccac

     1801 attccctttt tagtaaaata ttcagaaata atttaaatac atcattgcaa tgaaaataaa

     1861 tgttttttat taggcagaat ccagatgctc aaggcccttc ataatatccc ccagtttagt

     1921 agttggactt agggaacaaa ggaaccttta atagaaattg gacagcaaga aagcgagctt

     1981 ctagcttatc ctcagtcctg ctcctctgcc acaaagtgca cgcagttgcc ggccgggtcg

     2041 cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag

     2101 gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg

     2161 cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg

     2221 aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag

     2281 aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc

     2341 ggaacggcac tggtcaactt ggccatgatg gctcctcctg tcaggagagg aaagagaaga

     2401 aggttagtac aattgctata gtgagttgta ttatactatg cagatatact atgccaatga

     2461 ttaattgtca aactagggct gcagggttca tagtgccact tttcctgcac tgccccatct

     2521 cctgcccacc ctttcccagg catagacagt cagtgactta ccaaactcac aggagggaga

     2581 aggcagaagc ttgagacaga cccgcgggac cgccgaactg cgaggggacg tggctagggc

     2641 ggcttctttt atggtgcgcc ggccctcgga ggcagggcgc tcggggaggc ctagcggcca

     2701 atctgcggtg gcaggaggcg gggccgaagg ccgtgcctga ccaatccgga gcacatagga

     2761 gtctcagccc cccgccccaa agcaagggga agtcacgcgc ctgtagcgcc agcgtgttgt

     2821 gaaatggggg cttggggggg ttggggccct gactagtcaa aacaaactcc cattgacgtc

     2881 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta

     2941 ctgccaaaac cgcatcatca tggtaatagc gatgactaat acgtagatgt actgccaagt

     3001 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt

     3061 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt

     3121 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg

     3181 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc

     3241 atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc aaaaggccag

     3301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc

     3361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta

     3421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg

     3481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc

     3541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac

     3601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac

     3661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg

     3721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga

     3781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt

     3841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag

     3901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct

     3961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag ttaattaaca

     4021 tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt

     4081 tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag

     4141 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat cgaa

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