LOCUS       sequence-456849-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a de novo designed, FRET-based 
            neoLux1.2-CyOFP1 fusion bioluminescent probe (max. Em=588).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  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 6422)
  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: pcDNA3-luxOFP
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     580..599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    884..1005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(921..940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     975..994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1054..1070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1054..1070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1067..1089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1078..1094
                     /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)."
     promoter        complement(join(1102..1108,1109..1126,1127..1132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1147..1168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1285..1302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1456..2044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1536..1555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2215..3006,3007..3075))
                     /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     2838..2857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3076..3180)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3255..3274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3446..3825
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3826..4029
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3980..4000
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4074..4093
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4074..4092
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5224..5241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5230..5454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5500..5928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5587..5606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5797..5818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5937..5957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5942..6271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6122..6257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6184..6203
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6338..6422,1..710)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6392..6411)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
       61 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
      121 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      181 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      241 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      301 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      361 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
      421 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
      481 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
      541 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
      601 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
      661 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
      721 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
      781 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
      841 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
      901 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      961 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1021 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1081 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1141 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1201 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1261 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1321 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1861 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2161 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2221 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2281 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2341 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2401 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2461 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2521 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2581 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2641 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2701 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2761 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2821 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2881 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2941 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3001 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3061 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3121 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3181 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3241 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3301 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3361 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     3421 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     3481 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3541 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3601 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3661 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3721 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3781 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3841 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3901 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3961 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4021 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     4081 actcactata gggagaccca agctggctag cgtttaaact taagcttgcc gccaccatga
     4141 ccgacgagga gatcgccgag ttcgtgaagg ccttctacga ggcccttgac gctggagacg
     4201 ctgagacagc tgctgacctg ctgttcggag ccggatgcaa ggagattcac ctgtgggacg
     4261 gcaccgtgtt caccagcaag gaaggcttcg agacatggtt caagaagctg cagagcgccg
     4321 tgaccggcgc caagagaaag atcgagagcg tggagatcaa ggacggcgag gccgtggtga
     4381 agctgaccct gactgctaca tacaaggcca ccggcaagaa gttcgtcgtg gagctggagc
     4441 acaccttcac cttcgacaga gagaggaacg agctggtcga ggtgaaggtg accgccaccc
     4501 cgtatatcaa ggagaacatg agaagcaagc tgtacctgga gggcagcgtg aacggccacc
     4561 agttcaagtg cactcacgaa ggcgaaggca agccttacga gggcaagcag accaacagaa
     4621 tcaaggtggt ggaaggcggc cccctgccct tcgccttcga catcttagct acacacttca
     4681 tgtacggcag caaggtgttc atcaagtacc ccgccgacct gcccgactac tttaagcagt
     4741 ctttccccga gggcttcacc tgggagagag tgatggtgtt cgaggacggt ggagtgctga
     4801 ccgccaccca agacacctct ctgcaagacg gtgagctcat ctacaacgtg aaagtgagag
     4861 gagtgaactt ccccgccaat ggccccgtga tgcagaagaa gaccctggga tgggaaccta
     4921 gcaccgagac aatgtaccct gccgacggcg gtctggaagg aagatgcgac aaggccctga
     4981 agttggtggg cggtggccac ctgcacgtca acttcaaaac cacatacaag agcaagaagc
     5041 ccgtgaaaat gccaggcgtg cactacgtgg acagacggct ggagagaatt aaggaagccg
     5101 acaacgagac atacgtcgag cagtacgagc acgccgtggc cagatacagc aaccttggcg
     5161 gaggcatgga cgaactgtac aagtaactcg agtctagagg gcccgtttaa acccgctgat
     5221 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5281 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5341 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5401 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5461 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     5521 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5581 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5641 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5701 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5761 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5821 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5881 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     5941 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6001 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6061 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6121 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6181 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     6241 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     6301 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     6361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6421 aa
//
