pET28a- EGFP, 2 ug

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    pET28a-EGFP

    Search name

    pET28a-EGFP,Plasmid pET28a-EGFP,pET28a-EGFP vector

    Bacterial Resistance:Kanamycin
    Growth Strain:DH5α
    Expression: Bacterial
    Use:pET Plasmid

    Promoter: T7/lac
    Replicator: ColE1, ori, F1, ori
    Terminator: T7 Terminator
    Plasmid classification: large intestine series plasmid, large intestine fluorescent plasmid, large intestine green plasmid
    Plasmid size: 6050bp
    Plasmid labels: N-6 * His, N-Thrombin, N-EGFP, C-6 * His
    Prokaryotic resistance: kanamycin Kan (50 g/ml)
    Clone strain: DH5 alpha and other Escherichia coli
    Culture conditions: 37 DEG C, aerobic, LB
    Expressing host: BL21 (DE3) and other E. coli
    Culture conditions: 37 DEG C, aerobic, LB
    Induction: IPTG or lactose and its analogues
    Primers for 5'sequencing: T7 (TAATACGACTCACTATAGGG)
    Primers for 3'sequencing: T7-ter (TGCTAGTTATTGCTCAGCGG)

    Sequence

    LOCUS       Exported                6050 bp ds-DNA     circular SYN 12-JUL-2017
    DEFINITION  synthetic circular DNA
    KEYWORDS    pET28a-EGFP
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 6050)
      TITLE     Direct Submission
      JOURNAL   Exported 2015-4-17 
    REFERENCE   2  (bases 1 to 6050)
      AUTHORS   .
      TITLE     Direct Submission
      JOURNAL   Exported Wednesday, July 12, 2017
    FEATURES             Location/Qualifiers
         source          1..6050
                         /organism="synthetic DNA construct"
                         /mol_type="other DNA"
         rep_origin      12..467
                         /direction=RIGHT
                         /note="f1 ori"
                         /note="f1 ori
                         f1 bacteriophage origin of replication; arrow indicates
                         direction of (+) strand synthesis"
         CDS             complement(560..1375)
                         /codon_start=1
                         /gene="aph(3')-Ia"
                         /product="aminoglycoside phosphotransferase"
                         /note="aph(3')-Ia"
                         /note="KanR
                         confers resistance to kanamycin in bacteria or G418
                         (Geneticin(R)) in eukaryotes"
                         /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                         KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                         TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                         SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                         ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
         rep_origin      1497..2085
                         /direction=RIGHT
                         /note="ori"
                         /note="ori
                         high-copy-number ColE1/pMB1/pBR322/pUC origin of
                         replication"
         misc_feature    2271..2413
                         /note="bom"
                         /note="bom
                         basis of mobility region from pBR322"
         CDS             complement(2515..2706)
                         /codon_start=1
                         /gene="rop"
                         /product="Rop protein, which maintains plasmids at low copy
                         number"
                         /note="rop"
                         /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                         DELYRSCLARFGDDGENL"
         CDS             complement(3515..4597)
                         /codon_start=1
                         /gene="lacI"
                         /product="lac repressor"
                         /note="lacI"
                         /note="lacI
                         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(4598..4675)
                         /gene="lacI"
                         /note="lacI lacI promoter"
                         /note="lacI promoter"
         promoter        4984..5002
                         /note="T7 promoter"
                         /note="T7 promoter
                         promoter for bacteriophage T7 RNA polymerase"
         CDS             5083..5100
                         /codon_start=1
                         /product="6xHis affinity tag"
                         /note="6xHis affinity tag"
                         /note="6xHis"
                         /translation="HHHHHH"
         CDS             5131..5847
                         /codon_start=1
                         /product="enhanced GFP"
                         /note="enhanced GFP"
                         /note="EGFP
                         mammalian codon-optimized"
                         /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                         KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                         GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                         VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                         EFVTAAGITLGMDELYK"
         misc_feature    5848..5893
                         /note="MCS"
         CDS             5894..5911
                         /codon_start=1
                         /product="6xHis affinity tag"
                         /note="6xHis affinity tag"
                         /note="6xHis"