LOCUS       sequence-456847-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a de novo designed, FRET-based 
            neoLux1.2-mGold fusion bioluminescent probe (max. Em=530nm).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  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 6437)
  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-luxGold
FEATURES             Location/Qualifiers
     source          1..6437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(860..877)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    866..1090
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1136..1564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1223..1242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1433..1454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1573..1593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1578..1907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1758..1893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1820..1839
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1974..2768
                     /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
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIALYRLLDEFF"
     primer_bind     complement(2028..2047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2638..2657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2942..3063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2979..2998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3033..3052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3112..3128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3112..3128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3125..3147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3136..3152
                     /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(3160..3166,3167..3184,3185..3190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3205..3226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3343..3360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3514..4102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3594..3613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4273..5064,5065..5133))
                     /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     4896..4915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5134..5238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5313..5332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5504..5883
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5884..6087
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6038..6058
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6132..6151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6132..6150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagctgaccc tgactgctac atacaaggcc accggcaaga agttcgtcgt ggagctggag
       61 cacaccttca ccttcgacag agagaggaac gagctggtcg aggtgaaggt gaccgccacc
      121 ccttacttta ccggcgtggt gcctatcctg gtggagcttg acggcgatgt gaacggccat
      181 aagttcagcg tgagcgggga gggagagggt gacgccacat acggtaagct gacactgaag
      241 ttcatctgca ccacaggcaa actgcctgtg ccctggccta ccctggttac tagcctggga
      301 tacggcctgc agtgcttcgc cagatacccc gaccacatga agcagcacga ctttttcaag
      361 agcgccatgc cagagggcta cgtgcaggag agaaccatct tcttcaagga cgatggaaat
      421 tacaagacca gagccgaggt gaaattcgag ggcgacaccc ttgtgaacag aatcgagctg
      481 aagggcatcg acttcaaaga agacggcaac attctgggcc acaagctgga atacaactac
      541 aactctcaca acgtgtacat caccgccgac aagcagaaga acgggattaa ggccaacttt
      601 aagatccgtc acaatatcga ggacggcggc gtgcagctgg ctgaccacta tcagcagaac
      661 acccctatcg gagacggccc cgtgctgctg cctgataacc actacttgag ctaccaaagc
      721 aagctgagca aagaccccaa cgagaagcgg gatcacatgg tgcttctgga gttcgtgacc
      781 gccgccggca tcacacttgg catggacgag ctgtacaagt aactcgagtc tagagggccc
      841 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
      901 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
      961 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1021 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1081 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1141 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1201 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1261 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1321 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1381 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1441 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1501 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1561 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     1621 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     1681 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     1741 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1801 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     1861 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     1921 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     1981 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2041 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2101 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2161 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2221 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2281 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2341 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2401 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2461 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     2521 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2581 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     2641 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     2701 tttacggtat cgccgctccc gattcgcagc gcatcgcctt atatcgcctt cttgacgagt
     2761 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     2821 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     2881 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     2941 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3001 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3061 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3241 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5221 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     5281 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5341 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     5401 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     5461 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     5521 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5581 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5641 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5701 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5761 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5821 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5881 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5941 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     6001 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     6061 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     6121 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
     6181 agcttgccgc caccatgacc gacgaggaga tcgccgagtt cgtgaaggcc ttctacgagg
     6241 cccttgacgc tggagacgct gagacagctg ctgacctgct gttcggagcc ggatgcaagg
     6301 agattcacct gtgggacggc accgtgttca ccagcaagga aggcttcgag acatggttca
     6361 agaagctgca gagcgccgtg accggcgcca agagaaagat cgagagcgtg gagatcaagg
     6421 acggcgaggc cgtggtg
//
