LOCUS       sequence-388986-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BRET lux luciferase with Venus for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  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 5810)
  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:Venus+luxA/pRSETB Duet
FEATURES             Location/Qualifiers
     source          1..5810
                     /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"
     CDS             join(1520..1522,1523..1525,1526..2239)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1565..1586)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1826..1845)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2173..2194
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2305..2323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2324..2348
                     /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             2391..3473
                     /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(3545..3563)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3559..3606
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3703..4158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4000..4021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4184..4288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4289..4357,4358..5149)
                     /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(4507..4526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5320..5810,1..98)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(5809..5810,1..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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1561 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1621 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     1681 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg
     1741 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1801 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1861 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1921 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1981 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2041 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2101 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     2161 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2221 ggacgagctg tacaagtaag cggccgcata atgcttaagt cgaacagaaa gtaatcgtat
     2281 tgtacacggc cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata
     2341 acaattcccc atcttagtat attagttaag tataagaagg agatatacat atgaaatttg
     2401 gaaacttttt gcttacatac caacctcccc aattttctca aacagaggta atgaaacgtt
     2461 tggttaaatt aggtcgcatc tctgaggagt gtggttttga taccgtatgg ttactggagc
     2521 atcatttcac ggagtttggt ttgcttggta acccttatgt cgctgctgca tatttacttg
     2581 gcgcgactaa aaaattgaat gtaggaactg ccgctattgt tcttcccaca gcccatccag
     2641 tacgccaact tgaagatgtg aatttattgg atcaaatgtc aaaaggacga tttcggtttg
     2701 gtatttgccg agggctttac aacaaggact ttcgcgtatt cggcacagat atgaataaca
     2761 gtcgcgcctt agcggaatgc tggtacgggc tgataaagaa tggcatgaca gagggatata
     2821 tggaagctga taatgaacat atcaagttcc ataaggtaaa agtaaacccc gcggcgtata
     2881 gcagaggtgg cgcaccggtt tatgtggtgg ctgaatcagc ttcgacgact gagtgggctg
     2941 ctcaatttgg cctaccgatg atattaagtt ggattataaa tactaacgaa aagaaagcac
     3001 aacttgagct ttataatgaa gtggctcaag aatatgggca cgatattcat aatatcgacc
     3061 attgcttatc atatataaca tctgtagatc atgactcaat taaagcgaaa gagatttgcc
     3121 ggaaatttct ggggcattgg tatgattctt atgtgaatgc tacgactatt tttgatgatt
     3181 cagaccaaac aagaggttat gatttcaata aagggcagtg gcgtgacttt gtattaaaag
     3241 gacataaaga tactaatcgc cgtattgatt acagttacga aatcaatccc gtgggaacgc
     3301 cgcaggaatg tattgacata attcaaaaag acattgatgc tacaggaata tcaaatattt
     3361 gttgtggatt tgaagctaat ggaacagtag acgaaattat tgcttccatg aagctcttcc
     3421 agtctgatgt catgccattt cttaaagaaa aacaacgttc gctattatat tagggtacca
     3481 tggaattcga agcttgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     3541 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     3601 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     3661 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3721 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3781 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3841 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3901 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3961 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4021 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4081 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4141 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
//
