LOCUS       Exported                6749 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLR4a, complete sequence.
ACCESSION   KU744688
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6749)
  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  JOURNAL   Unpublished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 6749)
  AUTHORS   Koksharov MI.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-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 6749)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6749
                     /organism="Cloning vector pLR4a"
                     /mol_type="other DNA"
                     /db_xref="taxon:1898663"
     CDS             complement(5..757)
                     /codon_start=1
                     /product="transcription activator"
                     /label=transcription activator
                     /note="LuxR"
                     /protein_id="AOS51030.1"
                     /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="AOS51031.1"
                     /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..3285
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /note="derived from Luciola mingrelica; contains 
                     carboxy-terminal His-tag; contains additional BstBI, SalI, 
                     and XhoI restriction sites; luc_Lmi"
                     /protein_id="AOS51032.1"
                     /translation="MEMEKEENVVYGPLPFYPIEEGSAGIQLHKYMHQYAKLGAIAFSN
                     ALTGVDISYQEYFDITCRLAEAMKNFGMKPEEHIALCSENCEEFFIPVLAGLYIGVAVA
                     PTNEIYTLRELNHSLGIAQPTIVFSSRKGLPKVLEVQKTVTCIKKIVILDSKVNFGGHD
                     CMETFIKKHVELGFQPSSFVPIDVKNRKQHVALLMNSSGSTGLPKGVRITHEGAVTRFS
                     HAKDPIYGNQVSPGTAILTVVPFHHGFGMFTTLGYFACGYRVVMLTKFDEELFLRTLQD
                     YKCTSVILVPTLFAILNKSELIDKFDLSNLTEIASGGAPLAKEVGEAVARRFNLPGVRQ
                     GYGLTETTSAFIITPEGDDKPGASGKVVPLFKVKVIDLDTKKTLGVNRRGEICVKGPSL
                     MLGYSNNPEATRETIDEEGWLHTGDIGYYDEDEHFFIVDRLKSLIKYKGYQVPPAELES
                     VLLQHPNIFDAGVAGVPDPDAGELPGAVVVMEKGKTMTEKEIVDYVNSQVVNHKRLRGG
                     VRFVDEVPKGLTGKIDAKVIREILKKPQSGPVEHHHHHH"
     CDS             3265..3282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3335..3353)
                     /label=T7 terminator reverse primer
                     /note="T7 terminator reverse primer"
     terminator      3349..3396
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      3350..3396
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     rep_origin      3433..3888
                     /label=f1 origin
                     /note="f1 origin"
     promoter        3915..4019
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3950..3978
                     /regulatory_class="promoter"
                     /note="bla promoter"
     CDS             4020..4880
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="AOS51033.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5035..5654
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      5051..5639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5825..5965
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6067..6258)
                     /codon_start=1
                     /product="repressor of primer"
                     /label=repressor of primer
                     /note="ROP"
                     /protein_id="AOS51034.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"