LOCUS       Exported                5497 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  EGFP expression vector pBE, complete sequence.
ACCESSION   AY237157
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5497)
  AUTHORS   Colas J-F., Schoenwolf GC.
  TITLE     Assessing the contribution of gene products to the form-shaping 
            events of neurulation: a transgenic approach in chick
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5497)
  AUTHORS   Colas J-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2003) Neurobiology and Anatomy, University of 
            Utah, 20 North 1900 East MREB401, Salt Lake City, UT 84132-3401, USA
REFERENCE   3  (bases 1 to 5497)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5497
                     /organism="EGFP expression vector pBE"
                     /mol_type="other DNA"
                     /note="expresses enhanced green fluorescent protein (EGFP) 
                     when electroporated into the ectoderm of early chick 
                     embryos at gastrulation and neurulation stages"
                     /db_xref="taxon:235454"
     misc_feature    53..1402
                     /note="NcoI-SalI fragment (beta-actin promoter) of the 
                     pMES vector; NcoI restriction site is truncated"
     regulatory      57..334
                     /regulatory_class="promoter"
                     /note="Gallus gallus cytoplasmic beta-actin gene, GenBank 
                     Accession Number X00182"
     promoter        57..334
                     /label=chicken beta-actin promoter
     intron          335..1344
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     intron          335..1295
                     /note="Gallus gallus cytoplasmic beta-actin gene, GenBank 
                     Accession Number X00182; intron or putative primary 
                     transcript"
     intron          1304..1344
                     /note="Oryctolagus cuniculus beta1-globin gene, GenBank 
                     Accession Number K03256; intron 2"
     exon            1345..1398
                     /note="Oryctolagus cuniculus beta1-globin gene, GenBank 
                     Accession Number K03256; exon 3"
     misc_feature    1403..52
                     /note="sequence from cloning vector pEGFP-1, GenBank 
                     Accession Number U55761"
     regulatory      1437..1446
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1443..2162
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2285..2406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2413..2868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2895..2999
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3001..3358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3209..3344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3393..4187
                     /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    4419..4466
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4795..5383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"