LOCUS       Exported                5118 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEGFP-C1-M, complete sequence.
ACCESSION   KY499147
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5118)
  AUTHORS   Zhang X, Wang XY, Jia YL, Guo X, Wang YF, Wang TY.
  TITLE     A Vector Based on the Chicken Hypersensitive Site 4 Insulator 
            Element Replicates Episomally in Mammalian Cell
  JOURNAL   Curr Gene Ther (2017) In press
  PUBMED    28155604
REFERENCE   2  (bases 1 to 5118)
  AUTHORS   Wang T.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2017) Department of Biochemistry and Molecular 
            Biology, Xinxiang Medical University, Jinsui Road, Xinxiang, Henan 
            453000, Xinxiang
REFERENCE   3  (bases 1 to 5118)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5118
                     /organism="Cloning vector pEGFP-C1-M"
                     /mol_type="other DNA"
                     /label=extrachromosomal
                     /note="extrachromosomal"
                     /db_xref="taxon:1961142"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1717..1782
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1906..2027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2034..2489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2516..2620
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2622..2979
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2830..2965
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3014..3808
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4040..4087
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4416..5004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"