LOCUS       Exported                6802 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Inducible protein expression vector pReg Neo, complete sequence.
ACCESSION   EF030522
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6802)
  AUTHORS   Vidugiriene J, Schenborn E, McNamara B.
  TITLE     Regulated Mammalian Expression System Technical Manual, TM289
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6802)
  AUTHORS   Vidugiriene J, Schenborn E, McNamara B.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2006) Scientific Communications, Promega 
            Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 6802)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6802
                     /organism="Inducible protein expression vector pReg Neo"
                     /mol_type="other DNA"
                     /db_xref="taxon:410004"
     promoter        95..113
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      141..433
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     misc_feature    444..587
                     /label=6X lambda operator
                     /note="6X lambda operator"
     regulatory      600..659
                     /regulatory_class="promoter"
                     /note="minimal CMV promoter"
     promoter        725..743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             770..2443
                     /codon_start=1
                     /gene="Rep-GyrB-AD"
                     /product="Rep/GyrB/AD fusion protein"
                     /label=Rep-GyrB-AD
                     /note="chimeric transactivator protein; chimera contiaining
                     Lambda repressor (DNA binding domain), GyrB (amino terminal
                     domain) and p65 activation domain (AD)"
                     /protein_id="ABK40095.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVGMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVDISNSYDSSSIKVLKGLDAVRKRPGMYIGDTD
                     DGTGLHHMVFEVVDNAIDEALAGHCKEIIVTIHADNSVSVQDDGRGIPTGIHPEEGVSA
                     AEVIMTVLHAGGKFDDNSYKVSGGLHGVGVSVVNALSQKLELVIQREGKIHRQIYEHGV
                     PQAPLAVTGETEKTGTMVRFWPSLETFTNVTEFEYEILAKRLRELSFLNSGVSIRLRDK
                     RDGKEDHFHYEGGPCMGPKKKRKVAIDQLTMVFPSGQISNQALALAPSSAPVLAQTMVP
                     SSAMVPLAQPPAPAPVLTPGPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALL
                     GNSTDPGVFTDLASVDNSEFQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDP
                     APTPLGTSGLPNGLSGDEDFSSIADMDFSALLSQISSSGQ"
     gene            770..2443
                     /gene="Rep-GyrB-AD"
                     /label=Rep-GyrB-AD
     CDS             1841..1861
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1889..2431
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     promoter        complement(2471..2488)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2497..2718
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     polyA_signal    2506..2627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2813..3268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      3330..3748
                     /regulatory_class="enhancer"
                     /note="SV40 early enhancer/promoter region"
     promoter        3385..3742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3593..3728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3793..4587
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /function="neomycin resistance"
                     /label=neomycin phosphotransferase
                     /protein_id="ABK40096.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      4651..4699
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyA signal"
     polyA_signal    4651..4699
                     /note="synthetic polyadenylation signal"
     promoter        5005..5109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5110..5970
                     /codon_start=1
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=beta-lactamase
                     /protein_id="ABK40097.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6141..6729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6695..6732
                     /label=ColE1-derived origin of replication
                     /note="ColE1-derived origin of replication"