LOCUS       Exported                7605 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBIND-ER (alpha), complete sequence.
ACCESSION   GQ229579
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  AUTHORS   Kopish K, Pagiuo A, TenHarmsel A.
  TITLE     GloResponse(TM) 9XGAL4UAS-luc2P HEK293 Cell Line Technical Bulletin,
            TB552
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7605)
  AUTHORS   Kopish K, Pagiuo A, TenHarmsel A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2009) Scientific Communications, Promega 
            Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 7605)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7605
                     /organism="Cloning vector pBIND-ER (alpha)"
                     /mol_type="other DNA"
                     /db_xref="taxon:659258"
     regulatory      1..742
                     /regulatory_class="promoter"
                     /note="CMV immediate-early enhancer/promoter region"
     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"
     CDS             1083..1520
                     /codon_start=1
                     /gene="Gal4"
                     /product="Gal4 DNA binding domain"
                     /label=Gal4
                     /note="Gal4 DBD; DNA binding domain"
                     /protein_id="ACT53144.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTV"
     gene            1083..1520
                     /gene="Gal4"
                     /label=Gal4
     misc_feature    1521..1553
                     /label=IgA linker
                     /note="IgA linker"
     CDS             1563..2456
                     /codon_start=1
                     /gene="ER-LBD"
                     /product="ER-LBD"
                     /label=ER-LBD
                     /note="estrogen receptor ligand binding domain"
                     /protein_id="ACT53143.1"
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVECMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     gene            1563..2456
                     /gene="ER-LBD"
                     /label=ER-LBD
     regulatory      2616..2837
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     polyA_signal    2625..2746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2936..3354
                     /regulatory_class="promoter"
                     /note="SV40 enhancer/early promoter"
     promoter        2991..3348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3199..3334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3393..3402
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3399..5174
                     /codon_start=1
                     /gene="hRluc-neo"
                     /product="hRluc-neo fusion protein"
                     /label=hRluc-neo
                     /note="renilla luciferase-neomycin phosphotransferase 
                     fusion protein"
                     /protein_id="ACT53145.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQTGGGSGGGGSGGGGSGGIEQDGLHAGSPA
                     AWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLAT
                     TGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATC
                     PFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGD
                     ACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAP
                     DSQRIAFYRLLDEFF"
     gene            3399..5174
                     /gene="hRluc-neo"
                     /label=hRluc-neo
     CDS             3399..4331
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4383..5174
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      5238..5556
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyadenylation signal"
     polyA_signal    5238..5286
                     /note="synthetic polyadenylation signal"
     promoter        5442..5546
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5547..6407
                     /codon_start=1
                     /gene="Amp(R)"
                     /product="beta-lactamase"
                     /label=Amp(R)
                     /note="resistance to ampicillin"
                     /protein_id="ACT53142.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5547..6407
                     /gene="Amp(R)"
                     /label=Amp(R)
     rep_origin      6562..6598
                     /label=ColE1-derived plasmid origin of replication
                     /note="ColE1-derived plasmid origin of replication"
     rep_origin      complement(6565..7153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7269..7554
                     /label=cer
                     /note="cer"
     misc_feature    7269..7552
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"