LOCUS       Exported                6743 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLR6-Ppy-WT, complete sequence.
ACCESSION   KU747014
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  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  JOURNAL   Unpublished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 6743)
  AUTHORS   Koksharov MI.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2016) Department of Chemistry, Lomonosov Moscow 
            State University, Leninskie gory, Bld. 1/11, Moscow, Moscow 119991, 
            Russia
REFERENCE   3  (bases 1 to 6743)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6743
                     /organism="Cloning vector pLR6-Ppy-WT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1903068"
     CDS             complement(5..757)
                     /codon_start=1
                     /product="LuxR"
                     /label=LuxR
                     /note="derived from Aliivibrio fischeri bacterial activator
                     of transcription"
                     /protein_id="AOR57254.1"
                     /translation="MKDINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKSSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSKRTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKS"
     CDS             976..1557
                     /codon_start=1
                     /product="LuxI"
                     /label=LuxI
                     /note="derived from Aliivibrio fischeri 
                     acyl-homoserine-lactone synthase; produces the autoinducer"
                     /protein_id="AOR57255.1"
                     /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..3279
                     /codon_start=1
                     /product="Ppy_Luc"
                     /label=Ppy_Luc
                     /note="derived from Photinus pyralis firefly luciferase (WT
                     version) from INSD accession M15077; contains C-terminal 
                     histidine tail"
                     /protein_id="AOR57258.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     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 binding region
                     /note="T7 terminator reverse primer binding region"
     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="Bla; ampicillin resistance protein"
                     /protein_id="AOR57256.1"
                     /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..5959
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6061..6252)
                     /codon_start=1
                     /product="repressor of primer"
                     /label=repressor of primer
                     /note="regulatory protein ROP"
                     /protein_id="AOR57257.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"