LOCUS       sequence-456842-        5041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a de novo designed, FRET-based 
            neoLux1.2-mKate2 fusion bioluminescent probe (max. Em=638).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5041)
  AUTHORS   Chen JY-H, Shi Q, Peng X, Habimana JdD, Wang J, Sobolewski W, Yeh 
            AH-W
  TITLE     De novo luciferases enable multiplexed bioluminescence imaging.
  JOURNAL   Chem. 2025 Mar 13;11(3):102346. doi: 10.1016/j.chempr.2024.10.013. 
            Epub 2024 Nov 12.
  PUBMED    40417169
REFERENCE   2  (bases 1 to 5041)
  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: "Chem."; 
            date: "2025-03-13"; volume: "11(3):102346. doi"; pages: " 
            10.1016/j.chempr.2024.10.013. Epub 2024 Nov 12"
COMMENT     Sequence Label: pBAD-luxKate
FEATURES             Location/Qualifiers
     source          1..5041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..188
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(229..246)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      399..485
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      577..604
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        623..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2520..2660
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2586..2605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2746..2768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2924..3802)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3829..4113
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4036..4055
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 cttatcgttc gaaaaagccg gcgaaaaact taaaaatgcc gggcgtgtat tatgtggatc
       61 gccgtctgga acgcataaaa gaagcggata aagaaaccta tgtcgaacag catgaagtgg
      121 cggtggcgcg ctattgcgat ctgccaagca aactgggcca tcgcggttct catcatcatc
      181 atcatcatta aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
      241 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
      301 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
      361 ggagtaagcc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      421 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      481 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      541 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
      601 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg tagtcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     2581 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     2641 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2701 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2761 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
     2821 aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     2881 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
     2941 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
     3001 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
     3061 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct
     3121 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
     3181 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
     3241 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt
     3301 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
     3361 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
     3421 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
     3481 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
     3541 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
     3601 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
     3661 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
     3721 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
     3781 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt
     3841 gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca
     3901 aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa
     3961 aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg
     4021 gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac
     4081 gctttttatc gcaactctct actgtttctc catacccgtt ttttgggcta acaggaggaa
     4141 ttaaccatgg gctcaggaac agacgaagaa atagcagaat tcgtaaaagc attctacgaa
     4201 gcactcgacg caggagacgc agaaacagca gcagacctac tattcggagc aggatgcaaa
     4261 gaaatacacc tatgggacgg aacagtattc acaagcaaag aaggattcga aacatggttc
     4321 aaaaaactac aaagcgcagt aacaggagca aaaagaaaaa tagaaagcgt agaaataaaa
     4381 gacggagaag cagtagtaaa actaacacta acagcaacat acaaagcaac aggaaaaaaa
     4441 ttcgtagtag aactagaaca cacattcaca ttcgacagag aaagaaacga actagtagaa
     4501 gtgaaagtca cagcaacacc atatattaaa gaaaatatgc acatgaaact gtatatggaa
     4561 ggcacggtga acaaccacca ttttaaatgt accagcgaag gtgaaggcaa accgtatgaa
     4621 ggtacgcaga ccatgcgcat taaagcggtt gaaggcggtc cgctgccgtt tgcgtttgat
     4681 attctggcga ccagctttat gtatggcagc aaaaccttta ttaaccatac ccagggcatt
     4741 ccggatttct ttaaacagag ctttccggaa ggttttacct gggaacgcgt gaccacgtac
     4801 gaagatggtg gcgtgctgac tgcgactcaa gataccagcc tgcaggatgg ctgcctgatt
     4861 tataacgtga aaattcgcgg tgtgaacttt ccgagcaacg gcccggtgat gcagaaaaag
     4921 accttaggct gggaagcgag caccgaaacc ctgtatccgg cggatggcgg tttagaaggc
     4981 cgtgcggata tggcgctgaa attagtgggc ggcggccatc tgatttgcaa cctgaaaacc
     5041 a
//
