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pET22-EGFP大肠杆菌绿色荧光分泌型表达载体pET22b-EGFP-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

  • 价  格:¥9852
  • 货  号:pET22-EGFP大肠杆菌绿色荧光分泌型表达载体pET22b-EGFP
  • 产  地:北京
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pET22-EGFP大肠杆菌绿色荧光分泌型表达载体

Alt Name: pET22b-EGFP plasmid

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心www.biovector.net


启动子:T7/lac
复制子:ColE1 ori,F1 ori
终止子:T7 terminator
质粒分类:大肠杆菌载体;PET系列表达质粒
质粒大小:6232bp
质粒标签:N-pelB,EGFP
原核抗性:氨苄青霉素Amp
克隆菌株:DH5α
培养条件:37℃,有氧,LB
表达宿主:BL21(DE3)
诱导方式:IPTG或乳糖及其类似物
5'测序引物:T7:TAATACGACTCACTATAGGG
3'测序引物:T7-ter:TGCTAGTTATTGCTCAGCGG
备注:分泌表达


LOCUS       Exported                6232 bp ds-DNA     circular SYN 01-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pET22b-EGFP
SOURCE      synthetic DNA construct
 ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
 AUTHORS   .
 TITLE     Direct Submission
 JOURNAL   Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
           http://www.biovector.net
FEATURES             Location/Qualifiers
    source          1..6232
                    /organism="synthetic DNA construct"
                    /mol_type="other DNA"
    rep_origin      12..467
                    /direction=RIGHT
                    /note="f1 ori"
                    /note="f1 bacteriophage origin of replication; arrow
                    indicates direction of (+) strand synthesis"
    promoter        494..598
                    /gene="bla"
                    /note="AmpR promoter"
    CDS             599..1459
                    /codon_start=1
                    /gene="bla"
                    /product="beta-lactamase"
                    /note="AmpR"
                    /note="confers resistance to ampicillin, carbenicillin, and
                    related antibiotics"
                    /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                    ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                    PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                    EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                    LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                    LIKHW"
    rep_origin      1630..2218
                    /direction=RIGHT
                    /note="ori"
                    /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                    replication"
    misc_feature    2404..2546
                    /note="bom"
                    /note="basis of mobility region from pBR322"
    CDS             complement(2648..2839)
                    /codon_start=1
                    /gene="rop"
                    /product="Rop protein, which maintains plasmids at low copy
                    number"
                    /note="rop"
                    /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                    DELYRSCLARFGDDGENL"
    CDS             complement(3648..4730)
                    /codon_start=1
                    /gene="lacI"
                    /product="lac repressor"
                    /note="lacI"
                    /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)."
                    /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                    NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                    EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                    EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                    MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                    YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                    ALADSLMQLARQVSRLESGQ"
    promoter        complement(4731..4808)
                    /gene="lacI"
                    /note="lacI promoter"
    promoter        5117..5135
                    /note="T7 promoter"
                    /note="promoter for bacteriophage T7 RNA polymerase"
    protein_bind    5136..5160
                    /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)."
    RBS             5175..5197
                    /note="efficient ribosome binding site from bacteriophage
                    T7 gene 10 (Olins and Rangwala, 1989)"
    CDS             5205..5270
                    /codon_start=1
                    /gene="pelB (fragment)"
                    /product="leader peptide for secretion"
                    /note="pelB signal sequence"
                    /translation="MKYLLPTAAAGLLLLAAQPAMA"
    CDS             5303..6022
                    /codon_start=1
                    /product="enhanced GFP"
                    /note="EGFP"
                    /note="mammalian codon-optimized"
                    /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                    KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                    GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                    VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                    EFVTAAGITLGMDELYK"
    CDS             6076..6093
                    /codon_start=1
                    /product="6xHis affinity tag"
                    /note="6xHis"
                    /translation="HHHHHH"
    terminator      6160..6207
                    /note="T7 terminator"
                    /note="transcription terminator for bacteriophage T7 RNA
                    polymerase"
ORIGIN
       1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
     481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
    1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
    2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
    2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
    2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
    2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
    2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
    2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
    2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
    2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
    2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
    3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
    3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
    3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
    3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
    3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
    3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
    3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
    3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
    3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
    3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
    3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
    3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
    3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
    3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
    3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
    3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
    4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
    4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
    4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
    4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
    4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
    4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
    4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
    4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
    4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
    4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
    4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
    4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
    4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
    4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
    4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
    4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
    4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
    5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
    5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
    5161 cctctagaaa taattttgtt taactttaag aaggagatat acatatgaaa tacctgctgc
    5221 cgaccgctgc tgctggtctg ctgctcctcg ctgcccagcc ggcgatggcc atggatatcg
    5281 gaattaattc ggatccgaat tcatggtgag caagggcgag gagctgttca ccggggtggt
    5341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
    5401 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
    5461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
    5521 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
    5581 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
    5641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
    5701 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
    5761 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
    5821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
    5881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
    5941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
    6001 catggacgag ctgtacaagt aagcggccgc tcgagtctag agggccctat tctaaagctt
    6061 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
    6121 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
    6181 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at
//


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