LOCUS       Exported                4810 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHis-GFP, complete sequence.
ACCESSION   AF525449
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4810)
  AUTHORS   Yang Y, Ma J, Song Z, Wu M.
  TITLE     HIV-1 TAT-mediated protein transduction and subcellular localization
            using novel expression vectors
  JOURNAL   FEBS Lett. 532 (1-2), 36-44 (2002)
  PUBMED    12459459
REFERENCE   2  (bases 1 to 4810)
  AUTHORS   Yang YH, Wu M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2002) Department of Molecular and Cellular 
            Biology, School of Life Sciences, University of Science and 
            Technology of China, 96 Jingzai Road, Hefei, Anhui 230026, P.R. 
            China
REFERENCE   3  (bases 1 to 4810)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4810
                     /organism="Expression vector pHis-GFP"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:206382"
     regulatory      1..589
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      59..465
                     /regulatory_class="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      554..560
                     /regulatory_class="TATA_box"
     regulatory      615..624
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    633..650
                     /label=histidine tag
                     /note="histidine tag"
     CDS             633..650
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    648..748
                     /note="multiple cloning site; MCS"
     misc_feature    660..677
                     /label=thrombin
                     /note="thrombin"
     CDS             660..677
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    690..742
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      750..759
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             756..1475
                     /codon_start=1
                     /product="enhanced green fluorescent protein"
                     /label=enhanced green fluorescent protein
                     /protein_id="AAO20041.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1598..1719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1629..1634
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 early signal"
     regulatory      1658..1663
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 early signal"
     rep_origin      1726..2181
                     /label=f1
                     /note="f1"
     promoter        2208..2312
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      2243..2248
                     /regulatory_class="minus_35_signal"
     regulatory      2266..2271
                     /regulatory_class="minus_10_signal"
     promoter        2314..2671
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      2355..2584
                     /regulatory_class="promoter"
                     /note="SV40 early"
     regulatory      2355..2426
                     /regulatory_class="enhancer"
                     /note="tandem repeat"
     regulatory      2427..2498
                     /regulatory_class="enhancer"
                     /note="tandem repeat"
     rep_origin      2522..2657
                     /label=SV40
                     /note="SV40"
     misc_feature    2578..2584
                     /label=SV40 early promoter element
                     /note="SV40 early promoter element"
     CDS             2706..3500
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /protein_id="AAO20042.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3732..3779
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      3736..3741
                     /regulatory_class="polyA_signal_sequence"
                     /note="herpes simplex virus (HSV) thymidine kinase signal"
     regulatory      3749..3754
                     /regulatory_class="polyA_signal_sequence"
                     /note="herpes simplex virus (HSV) thymidine kinase signal"
     rep_origin      4085..4728
                     /label=pUC
                     /note="pUC"
     rep_origin      4108..4696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"