LOCUS       Exported                4843 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHis-TAT-GFP, complete sequence.
ACCESSION   AF525447
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4843)
  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 4843)
  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 4843)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4843
                     /organism="Expression vector pHis-TAT-GFP"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:206380"
     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    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    684..716
                     /note="Region: synthetic HIV-1 TAT protein transduction 
                     domain (PTD)"
     misc_feature    717..781
                     /note="multiple cloning site; MCS"
     misc_feature    723..775
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      783..792
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             789..1508
                     /codon_start=1
                     /product="enhanced green fluorescent protein"
                     /label=enhanced green fluorescent protein
                     /protein_id="AAO20037.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1631..1752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1662..1667
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 early signal"
     regulatory      1691..1696
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 early signal"
     rep_origin      1759..2214
                     /label=f1
                     /note="f1"
     promoter        2241..2345
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      2276..2281
                     /regulatory_class="minus_35_signal"
     regulatory      2299..2304
                     /regulatory_class="minus_10_signal"
     promoter        2347..2704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      2388..2617
                     /regulatory_class="promoter"
                     /note="SV40 early"
     regulatory      2388..2459
                     /regulatory_class="enhancer"
                     /note="tandem repeat"
     regulatory      2460..2531
                     /regulatory_class="enhancer"
                     /note="tandem repeat"
     rep_origin      2555..2690
                     /label=SV40
                     /note="SV40"
     misc_feature    2611..2617
                     /label=SV40 early promoter element
                     /note="SV40 early promoter element"
     CDS             2739..3533
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /protein_id="AAO20038.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3765..3812
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      3769..3774
                     /regulatory_class="polyA_signal_sequence"
                     /note="herpes simplex virus (HSV) thymidine kinase signal"
     regulatory      3782..3787
                     /regulatory_class="polyA_signal_sequence"
                     /note="herpes simplex virus (HSV) thymidine kinase signal"
     rep_origin      complement(4118..4761)
                     /direction=LEFT
                     /label=pUC
                     /note="pUC"
     rep_origin      4141..4729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"