LOCUS       Exported                6143 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  HaloTag CMV-neo vector pHTN, complete sequence.
ACCESSION   JF920304
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6143)
  AUTHORS   Hartnett J, Slater M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAY-2011) Promega Corporation, 2800 Woods Hollow Road,
            Madison, WI 53711, USA
REFERENCE   2  (bases 1 to 6143)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6143
                     /organism="HaloTag CMV-neo vector pHTN"
                     /mol_type="other DNA"
                     /db_xref="taxon:1093974"
     regulatory      1..742
                     /regulatory_class="promoter"
                     /note="CMV immediate/early promoter"
     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 N-terminal tag
                     /note="HaloTag N-terminal tag"
     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 linker N terminus
                     /note="HaloTag linker N terminus"
     misc_feature    1970..1990
                     /label=TEV protease recognition site
                     /note="TEV protease recognition site"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     misc_feature    1997..2072
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      2135..2356
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     polyA_signal    2144..2265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2455..2873
                     /regulatory_class="promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        2510..2867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2718..2853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2771..2836
                     /label=SV40 minimum origin of replication
                     /note="SV40 minimum origin of replication"
     CDS             2918..3712
                     /codon_start=1
                     /gene="neoR"
                     /product="neomycin phosphotransferase"
                     /label=neoR
                     /protein_id="AEQ28650.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2918..3712
                     /gene="neoR"
                     /label=neoR
                     /note="antibiotic resistance protein"
     regulatory      3776..3824
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyadenylation signal"
     polyA_signal    3776..3824
                     /note="synthetic polyadenylation signal"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4085..4945
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEQ28651.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4085..4945
                     /gene="ampR"
                     /label=ampR
     rep_origin      5100..5136
                     /label=ColE1-derived origin of replication
                     /note="ColE1-derived origin of replication"
     rep_origin      complement(5103..5691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5807..6090
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"