LOCUS       Exported                5535 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Flexi vector pFN28K HaloTag(R) CMV-neo, complete sequence.
ACCESSION   JN129498
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5535)
  AUTHORS   McNamara B, Kennedy J.
  TITLE     pFN28K HaloTag(R) CMV-neo Flexi Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5535)
  AUTHORS   McNamara B, Kennedy J.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUN-2011) Scientific Communications, Promega 
            Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 5535)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5535
                     /organism="Flexi vector pFN28K HaloTag(R) CMV-neo"
                     /mol_type="other DNA"
                     /db_xref="taxon:1055496"
     regulatory      1..742
                     /regulatory_class="promoter"
                     /note="CMV immediate-early enhancer/promoter; EcolCR1 site 
                     removed"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          857..989
                     /note="chimeric intron"
     regulatory      1033..1052
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1067..1957
                     /label=HaloTag 7 N-terminal region
                     /note="HaloTag 7 N-terminal region"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    1958..1996
                     /label=HaloTag 7 linker
                     /note="HaloTag 7 linker"
     misc_feature    1970..1990
                     /label=TEV protease recognition sequence
                     /note="TEV protease recognition sequence"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     misc_feature    1997..2004
                     /note="SgfI site; used in Flexi vector cloning"
     CDS             2028..2363
                     /codon_start=1
                     /product="barnase"
                     /label=barnase
                     /protein_id="AEK70328.1"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    2365..2372
                     /note="PmeI site; used in Flexi vector cloning"
     regulatory      2524..2745
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     polyA_signal    2533..2654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2844..3262
                     /regulatory_class="promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2899..3256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3107..3242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3270..3336
                     /label=EM7 bacterial promoter
                     /note="EM7 bacterial promoter"
     promoter        3271..3318
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3350..4144
                     /codon_start=1
                     /gene="neoR"
                     /product="neomycin phosphotransferase"
                     /label=neoR
                     /note="G-418 resistance"
                     /protein_id="AEK70329.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3350..4144
                     /gene="neoR"
                     /label=neoR
     regulatory      4208..4256
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyadenylation signal"
     polyA_signal    4208..4256
                     /note="synthetic polyadenylation signal"
     rep_origin      4492..4528
                     /label=ColE1-derived plasmid origin of replication
                     /note="ColE1-derived plasmid origin of replication"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5199..5482
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"