LOCUS       Exported                7350 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pmirGLO, complete sequence.
ACCESSION   FJ376737
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  AUTHORS   Garvin D.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-OCT-2008) Scientific Communication, Promega 
            Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA
REFERENCE   2  (bases 1 to 7350)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7350
                     /organism="Cloning vector pmirGLO"
                     /mol_type="other DNA"
                     /db_xref="taxon:574161"
     regulatory      106..327
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late poly(A) signal"
     polyA_signal    115..236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      426..844
                     /regulatory_class="promoter"
                     /note="SV40 early enhancer/promoter"
     promoter        481..838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      689..824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      883..892
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             889..2664
                     /codon_start=1
                     /product="hRluc-neo fusion protein"
                     /label=hRluc-neo fusion protein
                     /note="Renilla luciferase-neomycin resistance fusion 
                     protein; used as a control reporter for normalization"
                     /protein_id="ACJ37464.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQTGGGSGGGGSGGGGSGGIEQDGLHAGSPA
                     AWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLAT
                     TGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATC
                     PFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGD
                     ACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAP
                     DSQRIAFYRLLDEFF"
     CDS             889..1821
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1873..2664
                     /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      2728..2776
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyadenylation signal"
     polyA_signal    2728..2776
                     /note="synthetic polyadenylation signal"
     promoter        2932..3036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3037..3897
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to ampicillin and is used a 
                     selectable marker in E. coli"
                     /protein_id="ACJ37465.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4052..4088
                     /label=ColE1-derived plasmid origin of replication
                     /note="ColE1-derived plasmid origin of replication"
     rep_origin      complement(4055..4643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4759..5042
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     regulatory      5094..5609
                     /regulatory_class="promoter"
                     /note="human phosphoglycerate kinase promoter"
     promoter        5099..5598
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      5639..5648
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5645..7297
                     /codon_start=1
                     /product="modified firefly luciferase"
                     /label=modified firefly luciferase
                     /note="luc2; reporter of miRNA activity in mammalian cells"
                     /protein_id="ACJ37466.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    7306..7350
                     /label=multiple cloning region
                     /note="multiple cloning region"