LOCUS       Exported                6731 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLR6-Eluc, complete sequence.
ACCESSION   KU756582
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6731)
  AUTHORS   Koksharov MI, Ugarova NN.
  TITLE     Fast and simple in vivo photographic assessment of luciferase 
            properties in bacterial cells to screen and compare mutants of 
            beetle luciferases
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6731)
  AUTHORS   Koksharov MI.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-FEB-2016) Department of Chemical Enzymology, Faculty 
            of Chemistry, Lomonosov Moscow State University, Leninskie gory, 
            Bld. 1/11, Moscow, Moscow 119991, Russia
REFERENCE   3  (bases 1 to 6731)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6731
                     /organism="Cloning vector pLR6-Eluc"
                     /mol_type="other DNA"
                     /db_xref="taxon:1890401"
     CDS             complement(5..757)
                     /codon_start=1
                     /product="LuxR"
                     /label=LuxR
                     /note="bacterial activator of transcription [Aliivibrio 
                     fischeri]"
                     /protein_id="AOC83871.1"
                     /translation="MKDINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKSSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSKRTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKS"
     CDS             976..1557
                     /codon_start=1
                     /product="LuxI"
                     /label=LuxI
                     /note="acyl-homoserine-lactone synthase [Aliivibrio 
                     fischeri]; produces the autoinducer"
                     /protein_id="AOC83872.1"
                     /translation="MTIMIKKSDFLAIPSEEYKGILSLRYQVFKQRLEWDLVVENNLES
                     DEYDNSNAEYIYACDDTENVSGCWRLLPTTGDYMLKSVFPELLGQQSAPKDPNIVELSR
                     FAVGKNSSKINNSASEITMKLFEAIYKHAVSQGITEYVTVTSTAIERFLKRIKVPCHRI
                     GDKEIHVLGDTKSVVLSMPINEQFKKAVLN"
     primer_bind     1519..1548
                     /label=F_end_luxI forward primer binding region
                     /note="F_end_luxI forward primer binding region"
     primer_bind     1584..1611
                     /label=F_start_pLRx forward primer binding region
                     /note="F_start_pLRx forward primer binding region"
     CDS             1615..3267
                     /codon_start=1
                     /product="Eluc"
                     /label=Eluc
                     /note="Pyrearinus termitilluminans beetle luciferase 
                     (Eluc version - human-codon optimized); modified 
                     C-terminus"
                     /protein_id="AOC83873.1"
                     /translation="MEREKNVVYGPEPKHPLGNFTAGEMLYNALHKHSHIPQAILDVMG
                     NESLSYQEFFDTTVKLGQSLQNCGYKMNDVVSICAENNKRFFIPIISAWYIGMVVAPVN
                     EDYIPDELCKVTGISKPILVFTTRKILPKVLEVKDRTNYIKRIIILDSEENLLGCESLH
                     NFMSRYSDNNLQTFKPLHYDPVDQVAAILCSSGTTGLPKGVMQTHRNICVRLTHASDPR
                     VGTQLIPGVSVLAYLPFFHAFGFSINLGYFMVGLRVVMLRRFNQEVFLKAIQDYEVRSV
                     INVPSTILFLSKSPLVDKYDLSTLAELCCGAAPLAKEVAEIAVKRLNLPGIRCGYGLTE
                     STSANIHTLHNEFKSGSLGKVTPYMAAKIIDRNTGEALGPNQVGELCIWGPMVTKGYVN
                     NPQATKEAIDDDGWLHSGDFGYYDEDEYFYIVDRYKELIKYKGYQVAPVELEEILLQHP
                     GIRDVAVVGIPDIEAGELPAGFVVKQPGAQLTAKEVYDFLAQRVSHSKYLRGGVRFVDS
                     IPRNVTGKISRKELREALMEKASGPLEHHHHHH"
     RBS             3087..3095
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             3247..3264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3317..3335)
                     /label=T7 terminator reverse primer binding region
                     /note="T7 terminator reverse primer binding region"
     terminator      3331..3378
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      3332..3378
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     rep_origin      3415..3870
                     /label=f1 origin
                     /note="f1 origin"
     promoter        3897..4001
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3932..3960
                     /regulatory_class="promoter"
                     /note="bla promoter"
     CDS             4002..4862
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance protein"
                     /protein_id="AOC83874.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5017..5636
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5033..5621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5807..5947
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6049..6240)
                     /codon_start=1
                     /product="ROP"
                     /label=ROP
                     /note="repressor of primer; regulatory protein"
                     /protein_id="AOC83875.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"