LOCUS       sequence-456851-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian 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 6383)
  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 6383)
  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-luxKOk
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1999..2018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2190..2569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2570..2773
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2724..2744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2818..2837
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2818..2836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3929..3946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3935..4159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4205..4633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4292..4311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4502..4523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4642..4662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4647..4976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4827..4962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4889..4908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5043..5837
                     /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(5097..5116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5707..5726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6011..6132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6048..6067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6102..6121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6181..6197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6181..6197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6194..6216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6205..6221
                     /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(6229..6235,6236..6253,6254..6259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6274..6295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     1981 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     2041 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     2101 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     2161 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     2221 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2281 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2341 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2401 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2461 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2521 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2581 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2641 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2701 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2761 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     2821 tacgactcac tatagggaga cccaagctgg ctagcgttta aacttaagct tgccgccacc
     2881 atgaccgacg aggagatcgc cgagttcgtg aaggccttct acgaggccct tgacgctgga
     2941 gacgctgaga cagctgctga cctgctgttc ggagccggat gcaaggagat tcacctgtgg
     3001 gacggcaccg tgttcaccag caaggaaggc ttcgagacat ggttcaagaa gctgcagagc
     3061 gccgtgaccg gcgccaagag aaagatcgag agcgtggaga tcaaggacgg cgaggccgtg
     3121 gtgaagctga ccctgactgc tacatacaag gccaccggca agaagttcgt cgtggagctg
     3181 gagcacacct tcaccttcga cagagagagg aacgagctgg tcgaggtgaa ggtgaccgcc
     3241 acccctatcc tgcccgagat gaagatgaga tactacatgg acggcagcgt gaacggccac
     3301 gagttcacca tcgagggaga agggacaggc agaccttacg agggccacca ggaaatgacc
     3361 ctgagagtga caatggccga gggcggtccc atgcccttcg ctttcgatct ggttagccac
     3421 gtgttctgct acgggcacag agtgttcaca aagtaccccg aggagatccc cgactacttc
     3481 aagcaggctt tccccgaagg cctgagctgg gagagaagcc tggagtttga ggacggagga
     3541 agcgctagcg tgtccgccca catcagcttg aggggcaaca ccttctacca caaaagcaag
     3601 ttcacaggcg ttaacttccc cgccgacggc cccatcatgc agaatcagag cgtggactgg
     3661 gaacctagca ccgagaagat caccgcaagc gacggcgtgc tgaagggcga tgtcacaatg
     3721 tacctgaagc tggaaggcgg cggcaaccac aagtgccagt tcaagaccac atacaaggcc
     3781 gccaaagaga tcctggagat gcccggcgac cactacatcg gccacagact ggtgcggaag
     3841 acagagggaa acatcaccga gcaggtggag gatgccgtgg cccacagcta actcgagtct
     3901 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3961 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4021 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4081 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4141 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4201 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4561 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4621 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4801 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4981 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5041 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5101 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5161 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5221 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5281 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5341 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5401 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5461 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5521 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5581 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5641 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5701 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5761 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5821 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5881 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5941 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6001 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6121 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6361 cggccaacgc gcggggagag gcg
//
