LOCUS       sequence-457161-        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"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1342..1359
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1400..1417)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1570..1656
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1748..1775
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1794..1885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1886..1954,1955..2746)
                     /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(2104..2123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2917..3505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3406..3425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3659..3676
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3691..3831
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3757..3776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3917..3939
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4095..4973)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gggctcagga acagacgaag aaatagcaga attcgtaaaa
      361 gcattctacg aagcactcga cgcaggagac gcagaaacag cagcagacct actattcgga
      421 gcaggatgca aagaaataca cctatgggac ggaacagtat tcacaagcaa agaaggattc
      481 gaaacatggt tcaaaaaact acaaagcgca gtaacaggag caaaaagaaa aatagaaagc
      541 gtagaaataa aagacggaga agcagtagta aaactaacac taacagcaac atacaaagca
      601 acaggaaaaa aattcgtagt agaactagaa cacacattca cattcgacag agaaagaaac
      661 gaactagtag aagtgaaagt cacagcaaca ccaattctgc cggaaatgaa aatgcgctat
      721 tatatggatg gtagcgtgaa cggccatgag tttaccattg aaggcgaagg caccggccgc
      781 ccgtatgaag gccatcaaga aatgaccctg cgtgtgacga tggcggaagg cggcccaatg
      841 ccatttgcgt ttgatctggt gagccatgtg ttttgctatg gccatcgcgt gtttacgaaa
      901 tatccggaag agattccgga ttattttaaa caggcgtttc ctgaaggtct gagctgggaa
      961 agaagcctgg agttcgaaga tggtggcagc gcgagcgtga gcgcacatat tagcctgcgg
     1021 ggtaatacct tctatcataa aagcaaattt accggcgtta actttccggc ggatggtccg
     1081 attatgcaga accagagcgt tgattgggaa ccgagcacgg agaaaattac cgccagcgat
     1141 ggtgtgctga aaggtgatgt gaccatgtat ctgaaactgg agggcggcgg caaccataaa
     1201 tgccaattta aaaccacgta caaagccgcg aaagaaatct tagaaatgcc gggcgaccat
     1261 tatattggcc atagactggt tcgcaaaacc gaaggtaaca ttaccgaaca ggttgaagat
     1321 gcggtggcgc atagcggttc tcatcatcat catcatcatt aaagcttggc tgttttggcg
     1381 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     1441 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     1501 aagtgaaacg ccgtagcgcc gatggtagtg tggagtaagc ccatgcgaga gtagggaact
     1561 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1621 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     1681 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     1741 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     1801 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1861 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1921 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1981 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2041 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2101 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     2161 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2221 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2281 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2341 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2401 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2461 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2521 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2581 atctggagcc ggtgagcgtg gtagtcgcgg tatcattgca gcactggggc cagatggtaa
     2641 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2701 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2761 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2821 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3061 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3541 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3601 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3661 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3721 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3781 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3841 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3901 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3961 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt
     4021 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     4081 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     4141 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     4201 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     4261 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     4321 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     4381 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     4441 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     4501 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     4561 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     4621 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     4681 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     4741 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     4801 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     4861 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     4921 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     4981 atactcccgc cattcagag
//
