LOCUS       sequence-456848-        4999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a de novo designed, FRET-based 
            neoLux1.2-mKOk fusion bioluminescent probe (max. Em=562nm).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  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 4999)
  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-luxKOk
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      25..52
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        71..162
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(163..231,232..1023)
                     /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(381..400)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1194..1782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1683..1702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1936..1953
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1968..2108
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2034..2053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2194..2216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2372..3250)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3277..3561
                     /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     3484..3503
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4618..4635
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4676..4693)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4846..4932
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
       61 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      121 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      181 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      241 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      301 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      361 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
      421 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      481 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      541 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      601 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      661 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      721 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      781 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      841 tggtttattg ctgataaatc tggagccggt gagcgtggta gtcgcggtat cattgcagca
      901 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      961 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1021 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1081 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1141 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1201 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1261 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1321 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1381 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1441 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1501 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1561 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1621 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1681 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1741 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1801 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1861 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1921 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     1981 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2041 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2101 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2161 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2221 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
     2281 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     2341 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     2401 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     2461 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     2521 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     2581 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     2641 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     2701 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     2761 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     2821 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     2881 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     2941 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     3001 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     3061 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     3121 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     3181 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     3241 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     3301 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     3361 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     3421 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     3481 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     3541 aactctctac tgtttctcca tacccgtttt ttgggctaac aggaggaatt aaccatgggc
     3601 tcaggaacag acgaagaaat agcagaattc gtaaaagcat tctacgaagc actcgacgca
     3661 ggagacgcag aaacagcagc agacctacta ttcggagcag gatgcaaaga aatacaccta
     3721 tgggacggaa cagtattcac aagcaaagaa ggattcgaaa catggttcaa aaaactacaa
     3781 agcgcagtaa caggagcaaa aagaaaaata gaaagcgtag aaataaaaga cggagaagca
     3841 gtagtaaaac taacactaac agcaacatac aaagcaacag gaaaaaaatt cgtagtagaa
     3901 ctagaacaca cattcacatt cgacagagaa agaaacgaac tagtagaagt gaaagtcaca
     3961 gcaacaccaa ttctgccgga aatgaaaatg cgctattata tggatggtag cgtgaacggc
     4021 catgagttta ccattgaagg cgaaggcacc ggccgcccgt atgaaggcca tcaagaaatg
     4081 accctgcgtg tgacgatggc ggaaggcggc ccaatgccat ttgcgtttga tctggtgagc
     4141 catgtgtttt gctatggcca tcgcgtgttt acgaaatatc cggaagagat tccggattat
     4201 tttaaacagg cgtttcctga aggtctgagc tgggaaagaa gcctggagtt cgaagatggt
     4261 ggcagcgcga gcgtgagcgc acatattagc ctgcggggta ataccttcta tcataaaagc
     4321 aaatttaccg gcgttaactt tccggcggat ggtccgatta tgcagaacca gagcgttgat
     4381 tgggaaccga gcacggagaa aattaccgcc agcgatggtg tgctgaaagg tgatgtgacc
     4441 atgtatctga aactggaggg cggcggcaac cataaatgcc aatttaaaac cacgtacaaa
     4501 gccgcgaaag aaatcttaga aatgccgggc gaccattata ttggccatag actggttcgc
     4561 aaaaccgaag gtaacattac cgaacaggtt gaagatgcgg tggcgcatag cggttctcat
     4621 catcatcatc atcattaaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     4681 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     4741 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     4801 gtagtgtgga gtaagcccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     4861 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     4921 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     4981 cgggcaggac gcccgccat
//
