LOCUS       Exported                6450 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Diatom vector pDEST-EGFP, complete sequence.
ACCESSION   LT727255
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6450)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6450)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6450
                     /organism="Diatom vector pDEST-EGFP"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4542."
                     /db_xref="taxon:1944721"
     regulatory      1..568
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     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
                     /note="unnamed protein product; EGFP"
                     /protein_id="SJL87896.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      complement(1402..1526)
                     /regulatory_class="attenuator"
                     /note="attR1"
     protein_bind    1402..1526
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1534..1586
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     promoter        1551..1581
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1635..2294
                     /codon_start=1
                     /note="unnamed protein product; chloramphenicol resistance"
                     /protein_id="SJL87897.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2636..2941
                     /codon_start=1
                     /note="unnamed protein product; ccdB"
                     /protein_id="SJL87898.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      2982..3106
                     /regulatory_class="attenuator"
                     /note="attR2"
     protein_bind    complement(2982..3106)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      complement(3216..3312)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3216..3312
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    3238..3359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3366..3821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3822..3917
                     /regulatory_class="promoter"
                     /note="ampicillin resistance promoter"
     promoter        3848..3952
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3918..3952
                     /label=5 UTR of amp, incl. RBS
                     /note="5 UTR of amp, incl. RBS"
     promoter        3954..4311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      3973..4254
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      4162..4297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4211..4286
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             4346..5140
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87899.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      5331..5400
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     polyA_signal    5372..5419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5748..6336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6118..6444
                     /label=pMB1 ori
                     /note="pMB1 ori"