LOCUS       sequence-388994-        5828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BRET lux luciferase with cp157Venus for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5828)
  AUTHORS   Kaku T, Sugiura K, Entani T, Osabe K, Nagai T
  TITLE     Enhanced brightness of bacterial luciferase by bioluminescence 
            resonance energy transfer.
  JOURNAL   Sci Rep. 2021 Jul 22;11(1):14994. doi: 10.1038/s41598-021-94551-4.
  PUBMED    34294849
REFERENCE   2  (bases 1 to 5828)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2021-07-22"; volume: "11(1):14994. doi"; pages: " 
            10.1038/s41598-021-94551-4"
COMMENT     Sequence Label: luxB:cp157Venus+luxA/pRSETB Duet
FEATURES             Location/Qualifiers
     source          1..5828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     252..269
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        354..372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             404..426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             446..463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             467..499
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(503..511,512..526)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             533..1513
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     primer_bind     1705..1726
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(1784..1786,1787..1789,1790..2248)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(1829..1850)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2090..2109)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        2323..2341
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2342..2366
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2409..3491
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     primer_bind     complement(3563..3581)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3577..3624
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3721..4176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3808..3827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4018..4039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4202..4306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4307..4375,4376..5167)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4525..4544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5338..5828,1..98)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(5827..5828,1..18)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
       61 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      121 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      181 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccaaa
      241 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      301 ctctccccgc gcgttggccg attcattaat gcaggatctc gatcccgcga aattaatacg
      361 actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga
      421 aggagatata catatgcggg gttctcatca tcatcatcat catggtatgg ctagcatgac
      481 tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc cgatgaaatt
      541 tggattgttc ttccttaact tcatcaattc aacaactgtt caagaacaaa gtatagttcg
      601 catgcaggaa ataacggagt atgttgataa gttgaatttt gaacagattt tagtgtatga
      661 aaatcatttt tcagataatg gtgttgtcgg cgctcctctg actgtttctg gttttctgct
      721 cggtttaaca gagaaaatta aaattggttc attaaatcac atcattacaa ctcatcatcc
      781 tgtccgcata gcggaggaag cttgcttatt ggatcagtta agtgaaggga gatttatttt
      841 agggtttagt gattgcgaaa aaaaagatga aatgcatttt tttaatcgcc cggttgaata
      901 tcaacagcaa ctatttgaag agtgttatga aatcattaac gatgctttaa caacaggcta
      961 ttgtaatcca gataacgatt tttatagctt ccctaaaata tctgtaaatc cccatgctta
     1021 tacgccaggc ggacctcgga aatatgtaac agcaaccagt catcatattg ttgagtgggc
     1081 ggccaaaaaa ggtattcctc tcatctttaa gtgggatgat tctaatgatg ttagatatga
     1141 atatgctgaa agatataaag ccgttgcgga taaatatgac gttgacctat cagagataga
     1201 ccatcagtta atgatattag ttaactataa cgaagatagt aataaagcta aacaagagac
     1261 gcgtgcattt attagtgatt atgttcttga aatgcaccct aatgaaaatt tcgaaaataa
     1321 acttgaagaa ataattgcag aaaacgctgt cggaaattat acggagtgta taactgcggc
     1381 taagttggca attgaaaagt gtggtgcgaa aagtgtattg ctgtcctttg aaccaatgaa
     1441 tgatttgatg agccaaaaaa atgtaatcaa tattgttgat gataatatta agaagtacca
     1501 catggaatat accgagctca tgcagaagaa cggcatcaag gccaacttca agatccgcca
     1561 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1621 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     1681 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1741 cactctcggc atggacgagc tgtacaaggg tggcagcggt ggcatggtga gcaagggcga
     1801 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1861 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1921 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     1981 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2041 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2101 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2161 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2221 caacagccac aacgtctata tcaccgccga caagtaagcg gccgcataat gcttaagtcg
     2281 aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag
     2341 gggaattgtg agcggataac aattccccat cttagtatat tagttaagta taagaaggag
     2401 atatacatat gaaatttgga aactttttgc ttacatacca acctccccaa ttttctcaaa
     2461 cagaggtaat gaaacgtttg gttaaattag gtcgcatctc tgaggagtgt ggttttgata
     2521 ccgtatggtt actggagcat catttcacgg agtttggttt gcttggtaac ccttatgtcg
     2581 ctgctgcata tttacttggc gcgactaaaa aattgaatgt aggaactgcc gctattgttc
     2641 ttcccacagc ccatccagta cgccaacttg aagatgtgaa tttattggat caaatgtcaa
     2701 aaggacgatt tcggtttggt atttgccgag ggctttacaa caaggacttt cgcgtattcg
     2761 gcacagatat gaataacagt cgcgccttag cggaatgctg gtacgggctg ataaagaatg
     2821 gcatgacaga gggatatatg gaagctgata atgaacatat caagttccat aaggtaaaag
     2881 taaaccccgc ggcgtatagc agaggtggcg caccggttta tgtggtggct gaatcagctt
     2941 cgacgactga gtgggctgct caatttggcc taccgatgat attaagttgg attataaata
     3001 ctaacgaaaa gaaagcacaa cttgagcttt ataatgaagt ggctcaagaa tatgggcacg
     3061 atattcataa tatcgaccat tgcttatcat atataacatc tgtagatcat gactcaatta
     3121 aagcgaaaga gatttgccgg aaatttctgg ggcattggta tgattcttat gtgaatgcta
     3181 cgactatttt tgatgattca gaccaaacaa gaggttatga tttcaataaa gggcagtggc
     3241 gtgactttgt attaaaagga cataaagata ctaatcgccg tattgattac agttacgaaa
     3301 tcaatcccgt gggaacgccg caggaatgta ttgacataat tcaaaaagac attgatgcta
     3361 caggaatatc aaatatttgt tgtggatttg aagctaatgg aacagtagac gaaattattg
     3421 cttccatgaa gctcttccag tctgatgtca tgccatttct taaagaaaaa caacgttcgc
     3481 tattatatta gggtaccatg gaattcgaag cttgatccgg ctgctaacaa agcccgaaag
     3541 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     3601 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc tggcgtaata
     3661 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     3721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3841 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4081 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4141 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4201 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4261 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4321 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4381 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4441 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4501 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4561 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4621 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4681 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4741 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4801 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4861 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4921 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4981 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5041 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5101 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5161 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5221 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5281 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5821 tccagggg
//
