LOCUS       Exported                6746 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLR6-Lno, complete sequence.
ACCESSION   KU756583
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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  JOURNAL   Unpublished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 6746)
  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 6746)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6746
                     /organism="Cloning vector pLR6-Lno"
                     /mol_type="other DNA"
                     /db_xref="taxon:1890677"
     CDS             complement(5..757)
                     /codon_start=1
                     /product="luxR"
                     /label=luxR
                     /note="bacterial activator of transcription [Aliivibrio 
                     fischeri]"
                     /protein_id="AOC83877.1"
                     /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="AOC83878.1"
                     /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..3282
                     /codon_start=1
                     /product="Lno_luc"
                     /EC_number="1.13.12.7"
                     /label=Lno_luc
                     /note="Lampyris noctiluca firefly luciferase; 
                     modifiedC-terminus"
                     /protein_id="AOC83876.1"
                     /translation="MEDAKNIMHGPAPFYPLEDGTAGEQLHKAMKRYAQVPGTIAFTDA
                     HAEVNITYSEYFEMACRLAETMKRYGLGLQHHIAVCSENSLQFFMPVCGALFIGVGVAP
                     TNDIYNERELYNSLSISQPTIVFCSKRALQKILGVQKKLPIIQKIVILDSREDYMGKQS
                     MYSFIESHLPAGFNEYDYIPDSFDRETATALIMNSSGSTGLPKGVELTHQNVCVRFSHC
                     RDPVFGNQIIPDTAILTVIPFHHGFGMFTTLGYLTCGFRIVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLVDKYDLSNLHEIASGGAPLAKEVGEAVAKRFKLPGIRQGY
                     GLTETTSAIIITPEGDDKPGACGKVVPFFSAKIVDLDTGKTLGVNQRGELCVKGPMIMK
                     GYVNNPEATSALIDKDGWLHSGDIAYYDKDGHFFIVDRLKSLIKYKGYQVPPAELESIL
                     LQHPFIFDAGVAGIPDPDAGELPAAVVVLEEGKTMTEQEVMDYVAGQVTASKRLRGGVK
                     FVDEVPKGLTGKIDGRKIREILMMGKKSGPVEHHHHHH"
     CDS             3262..3279
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3332..3350)
                     /label=T7 terminator reverse primer binding region
                     /note="T7 terminator reverse primer binding region"
     terminator      3346..3393
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      3347..3393
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     rep_origin      3430..3885
                     /label=f1 origin
                     /note="f1 origin"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3947..3975
                     /regulatory_class="promoter"
                     /note="bla promoter"
     CDS             4017..4877
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance protein"
                     /protein_id="AOC83879.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5032..5651
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5822..5962
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6064..6255)
                     /codon_start=1
                     /product="ROP"
                     /label=ROP
                     /note="repressor of primer; regulatory protein"
                     /protein_id="AOC83880.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"