LOCUS       sequence-388980-        7401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing luxA and luxB under dual T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  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 7401)
  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: luxA+luxB/pRSETB Duet
FEATURES             Location/Qualifiers
     source          1..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             26..43
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             47..79
                     /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(83..91,92..106)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             113..1195
                     /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"
     promoter        1261..1279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1280..1304
                     /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             1347..2330
                     /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     complement(2402..2420)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2416..2463
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2529..2909
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2616..2635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2826..2847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3099..3890,3891..3959))
                     /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     3722..3741
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3960..4031)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4972..4991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5132..5154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5148..5339)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6959..6978
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6998..7075)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7335..7353
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             join(7385..7401,1..6)
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 aggagatata catatgcggg gttctcatca tcatcatcat catggtatgg ctagcatgac
       61 tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc cgatgaaatt
      121 tggaaacttt ttgcttacat accaacctcc ccaattttct caaacagagg taatgaaacg
      181 tttggttaaa ttaggtcgca tctctgagga gtgtggtttt gataccgtat ggttactgga
      241 gcatcatttc acggagtttg gtttgcttgg taacccttat gtcgctgctg catatttact
      301 tggcgcgact aaaaaattga atgtaggaac tgccgctatt gttcttccca cagcccatcc
      361 agtacgccaa cttgaagatg tgaatttatt ggatcaaatg tcaaaaggac gatttcggtt
      421 tggtatttgc cgagggcttt acaacaagga ctttcgcgta ttcggcacag atatgaataa
      481 cagtcgcgcc ttagcggaat gctggtacgg gctgataaag aatggcatga cagagggata
      541 tatggaagct gataatgaac atatcaagtt ccataaggta aaagtaaacc ccgcggcgta
      601 tagcagaggt ggcgcaccgg tttatgtggt ggctgaatca gcttcgacga ctgagtgggc
      661 tgctcaattt ggcctaccga tgatattaag ttggattata aatactaacg aaaagaaagc
      721 acaacttgag ctttataatg aagtggctca agaatatggg cacgatattc ataatatcga
      781 ccattgctta tcatatataa catctgtaga tcatgactca attaaagcga aagagatttg
      841 ccggaaattt ctggggcatt ggtatgattc ttatgtgaat gctacgacta tttttgatga
      901 ttcagaccaa acaagaggtt atgatttcaa taaagggcag tggcgtgact ttgtattaaa
      961 aggacataaa gatactaatc gccgtattga ttacagttac gaaatcaatc ccgtgggaac
     1021 gccgcaggaa tgtattgaca taattcaaaa agacattgat gctacaggaa tatcaaatat
     1081 ttgttgtgga tttgaagcta atggaacagt agacgaaatt attgcttcca tgaagctctt
     1141 ccagtctgat gtcatgccat ttcttaaaga aaaacaacgt tcgctattat attaggcggc
     1201 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
     1261 taatacgact cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta
     1321 gttaagtata agaaggagat atacatatga aatttggatt gttcttcctt aacttcatca
     1381 attcaacaac tgttcaagaa caaagtatag ttcgcatgca ggaaataacg gagtatgttg
     1441 ataagttgaa ttttgaacag attttagtgt atgaaaatca tttttcagat aatggtgttg
     1501 tcggcgctcc tctgactgtt tctggttttc tgctcggttt aacagagaaa attaaaattg
     1561 gttcattaaa tcacatcatt acaactcatc atcctgtccg catagcggag gaagcttgct
     1621 tattggatca gttaagtgaa gggagattta ttttagggtt tagtgattgc gaaaaaaaag
     1681 atgaaatgca tttttttaat cgcccggttg aatatcaaca gcaactattt gaagagtgtt
     1741 atgaaatcat taacgatgct ttaacaacag gctattgtaa tccagataac gatttttata
     1801 gcttccctaa aatatctgta aatccccatg cttatacgcc aggcggacct cggaaatatg
     1861 taacagcaac cagtcatcat attgttgagt gggcggccaa aaaaggtatt cctctcatct
     1921 ttaagtggga tgattctaat gatgttagat atgaatatgc tgaaagatat aaagccgttg
     1981 cggataaata tgacgttgac ctatcagaga tagaccatca gttaatgata ttagttaact
     2041 ataacgaaga tagtaataaa gctaaacaag agacgcgtgc atttattagt gattatgttc
     2101 ttgaaatgca ccctaatgaa aatttcgaaa ataaacttga agaaataatt gcagaaaacg
     2161 ctgtcggaaa ttatacggag tgtataactg cggctaagtt ggcaattgaa aagtgtggtg
     2221 cgaaaagtgt attgctgtcc tttgaaccaa tgaatgattt gatgagccaa aaaaatgtaa
     2281 tcaatattgt tgatgataat attaagaagt accacatgga atatacctaa ggtaccatgg
     2341 aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     2401 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2461 ttgctgaaas gaggarsymy rymcsgatck sscktcccaa cagttgcgca gcctgaatgg
     2521 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2761 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2821 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2881 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2941 aaaatttaac gcgrrtytka msmwmakwkk aacgmwwact cacgttaagg gattttggtc
     3001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3061 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3121 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3181 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3241 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3301 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3361 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3421 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3481 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3541 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3601 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3661 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3721 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3781 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3841 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3901 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3961 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4021 atatttgaat gtakttaraa awmtwwwywa atakskswtg accaaaatcc cttaacgtga
     4081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4741 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4801 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4861 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgccyra trcgswawyy
     4921 kcyyctymcs smkckgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     4981 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     5041 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5101 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5161 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     5221 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     5281 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     5341 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     5401 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     5461 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     5521 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     5581 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     5641 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     5701 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     5761 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     5821 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     5881 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     5941 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     6001 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     6061 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     6121 tgatcgaagt katggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     6181 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     6241 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     6301 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     6361 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     6421 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     6481 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     6541 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     6601 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     6661 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccrcgc
     6721 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     6781 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     6841 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     6901 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     6961 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     7021 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     7081 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     7141 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     7201 ccgsccmccg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7261 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct
     7321 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
     7381 taattttgtt taactttaag a
//
