LOCUS       Exported                6745 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLR5-Ppy-luc2, complete sequence.
ACCESSION   KU744692
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Koksharov MI, Ugarova NN.
  TITLE     Fast and simple in vivo photographic assessment of luciferase 
            properties in E. coli cells to screen and compare mutants of beetle 
            luciferases
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6745)
  AUTHORS   Koksharov MI.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-FEB-2016) Department of Chemical Enzymology, Faculty 
            of Chemistry, Lomonosov Moscow State University, Leninskie gory, 
            Bld. 1/11, Moscow 119991, Russia
REFERENCE   3  (bases 1 to 6745)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6745
                     /organism="Cloning vector pLR5-Ppy-luc2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1898664"
     CDS             complement(5..757)
                     /codon_start=1
                     /product="transcription activator"
                     /label=transcription activator
                     /note="LuxR; derived from Aliivibrio fischeri"
                     /protein_id="AOS51050.1"
                     /translation="MKDINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKSSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSKRTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKS"
     CDS             976..1557
                     /codon_start=1
                     /product="acyl-homoserine-lactone synthase"
                     /label=acyl-homoserine-lactone synthase
                     /note="LuxI; derived from Aliivibrio fischeri; autoinducer"
                     /protein_id="AOS51051.1"
                     /translation="MTIMIKKSDFLAIPSEEYKGILSLRYQVFKQRLEWDLVVENNLES
                     DEYDNSNAEYIYACDDTENVSGCWRLLPTTGDYMLKSVFPELLGQQSAPKDPNIVELSR
                     FAVGKNSSKINNSASEITMKLFEAIYKHAVSQGITEYVTVTSTAIERFLKRIKVPCHRI
                     GDKEIHVLGDTKSVVLSMPINEQFKKAVLN"
     primer_bind     1519..1548
                     /label=F_end_luxI
                     /note="F_end_luxI"
     primer_bind     1584..1611
                     /label=F_start_pLRx
                     /note="F_start_pLRx"
     CDS             1615..3279
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /note="derived from Photinus pyralis; contains 
                     carboxy-terminal His-tag; Ppy_luc2"
                     /protein_id="AOS51054.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGPLEHHHHHH"
     CDS             3259..3276
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3329..3347)
                     /label=T7 terminator reverse primer
                     /note="T7 terminator reverse primer"
     terminator      3343..3390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      3344..3390
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     rep_origin      3427..3882
                     /label=f1 origin
                     /note="f1 origin"
     promoter        3909..4013
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3944..3972
                     /regulatory_class="promoter"
                     /note="bla promoter"
     CDS             4014..4874
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="AOS51052.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5029..5648
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5045..5633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5819..5961
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6063..6254)
                     /codon_start=1
                     /product="repressor of primer"
                     /label=repressor of primer
                     /note="ROP"
                     /protein_id="AOS51053.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"