LOCUS       Exported                4363 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pGL4.23-Lmi-PCre, complete sequence.
ACCESSION   KY025566
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4363)
  AUTHORS   Koksharov M.
  TITLE     Design of short-lived firefly luciferase transcriptional reporters 
            in mammalian cells: effects of N- or C-terminal modifications and 
            interspecies differences on reporter half-life
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4363)
  AUTHORS   Koksharov M, Ugarova N.
  TITLE     Fast and simple in vivo photographic assessment of luciferase 
            properties in bacterial cells to screen and compare mutant beetle 
            luciferases: novel mutants illuminate luciferase color determinants 
            and factors of protein stability in living cells
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4363)
  AUTHORS   Koksharov M.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-OCT-2016) Department of Chemical Enzymology, Faculty 
            of Chemistry, Lomonosov Moscow State University, Leninskie gory, 
            Bld. 1/11, Moscow, Moscow 119991, Russia
REFERENCE   4  (bases 1 to 4363)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4363
                     /organism="Reporter vector pGL4.23-Lmi-PCre"
                     /mol_type="other DNA"
                     /db_xref="taxon:1978307"
     misc_feature    1..144
                     /note="multiple cloning region with an inserted 
                     CRE-dependent promoter"
     regulatory      27..125
                     /regulatory_class="promoter"
                     /note="CREB-dependent promoter PCre (4x CREB binding 
                     sites)"
     protein_bind    44..51
                     /label=CREB binding site
                     /bound_moiety="CREB"
                     /note="CREB palindromic site"
     protein_bind    70..74
                     /label=CREB binding site
                     /bound_moiety="CREB"
                     /note="CREB half-site"
     protein_bind    80..86
                     /label=CREB binding site
                     /bound_moiety="CREB"
                     /note="CREB non-palindromic site"
     protein_bind    102..109
                     /label=CREB binding site
                     /bound_moiety="CREB"
                     /note="CREB palindromic site"
     promoter        151..182
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      152..182
                     /regulatory_class="promoter"
                     /note="minimal promoter"
     regulatory      209..218
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             215..1873
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /EC_number="1.13.12.7"
                     /label=luc
                     /note="Luciola mingrelica firefly luciferase (wild type)"
                     /protein_id="ARG47622.1"
                     /translation="MEMEKEENVVYGPLPFYPIEEGSAGIQLHKYMHQYAKLGAIAFSN
                     ALTGVDISYQEYFDITCRLAEAMKNFGMKPEEHIALCSENCEEFFIPVLAGLYIGVAVA
                     PTNEIYTLRELNHSLGIAQPTIVFSSRKGLPKVLEVQKTVTCIKKIVILDSKVNFGGHD
                     CMETFIKKHVELGFQPSSFVPIDVKNRKQHVALLMNSSGSTGLPKGVRITHEGAVTRFS
                     HAKDPIYGNQVSPGTAILTVVPFHHGFGMFTTLGYFACGYRVVMLTKFDEELFLRTLQD
                     YKCTSVILVPTLFAILNKSELIDKFDLSNLTEIASGGAPLAKEVGEAVARRFNLPGVRQ
                     GYGLTETTSAFIITPEGDDKPGASGKVVPLFKVKVIDLDTKKTLGVNRRGEICVKGPSL
                     MLGYSNNPEATRETIDEEGWLHTGDIGYYDEDEHFFIVDRLKSLIKYKGYQVPPAELES
                     VLLQHPNIFDAGVAGVPDPDAGELPGAVVVMEKGKTMTEKEIVDYVNSQVVNHKRLRGG
                     VRFVDEVPKGLTGKIDAKVIREILKKPQSGPLEAV"
     gene            215..1873
                     /gene="luc"
                     /label=luc
     regulatory      1908..2129
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late poly(A) signal"
     polyA_signal    1917..2038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2198..2216)
                     /label=RV4 reverse primer binding region
                     /note="RV4 reverse primer binding region"
     rep_origin      2442..3061
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(2457..3045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3245..4105)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ARG47623.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      4210..4363
                     /regulatory_class="polyA_signal_sequence"
                     /note="Synthetic poly(A) signal/transcriptional pause site"
     polyA_signal    4210..4258
                     /note="synthetic polyadenylation signal"
     misc_feature    4272..4363
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4312..4331
                     /label=RV3 forward primer binding region
                     /note="RV3 forward primer binding region"