LOCUS       sequence-456843-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a de novo designed, FRET-based 
            neoLux1.2-mNeonGreen fusion bioluminescent probe (max. Em=518nm).
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-luxNeon
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..71)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(225..813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(305..324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(984..1775,1776..1844))
                     /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     1607..1626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1845..1949)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2024..2043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2215..2594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2595..2798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2749..2769
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2843..2862
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2843..2861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3993..4010)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3999..4223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4269..4697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4356..4375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4566..4587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4706..4726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4711..5040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4891..5026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4953..4972
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5107..5901
                     /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(5161..5180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5771..5790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6075..6196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6112..6131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6166..6185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6245..6261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6245..6261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6258..6280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6269..6285
                     /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(6293..6299,6300..6317,6318..6323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6338..6359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
       61 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1981 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     2041 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     2101 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     2161 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     2221 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2281 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2341 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2401 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2461 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2521 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2581 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2641 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2701 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2761 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     2821 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
     2881 gtttaaactt aagcttgccg ccaccatgac cgacgaggag atcgccgagt tcgtgaaggc
     2941 cttctacgag gcccttgacg ctggagacgc tgagacagct gctgacctgc tgttcggagc
     3001 cggatgcaag gagattcacc tgtgggacgg caccgtgttc accagcaagg aaggcttcga
     3061 gacatggttc aagaagctgc agagcgccgt gaccggcgcc aagagaaaga tcgagagcgt
     3121 ggagatcaag gacggcgagg ccgtggtgaa gctgaccctg actgctacat acaaggccac
     3181 cggcaagaag ttcgtcgtgg agctggagca caccttcacc ttcgacagag agaggaacga
     3241 gctggtcgag gtgaaggtga ccgccacccc tctgaacagc ctgcccgcaa cacacgagct
     3301 gcacatcttc ggcagcatca acggcgtgga cttcgacatg gtgggccagg gcactggaaa
     3361 ccctaacgac ggctacgagg agctgaacct gaagtccacc aagggggacc ttcagttcag
     3421 cccctggatc ctggtgcccc acatcggata cggcttccac cagtacctgc cctaccctga
     3481 cggcatgagc cccttccaag ccgctatggt tgacggaagc ggctaccagg tgcacagaac
     3541 catgcagttc gaggacggcg ctagcctgac cgtgaactac agatacactt acgagggcag
     3601 ccacatcaaa ggcgaggccc aggtgaaggg cacaggattc cctgctgacg gacctgtgat
     3661 gaccaacagc ctcaccgccg ctgactggtg tagaagcaag aagacctacc ccaacgacaa
     3721 gaccatcatc agcaccttca agtggagcta caccaccggc aacggcaaga gatacagaag
     3781 caccgccaga acaacctaca ccttcgccaa gcctatggcc gccaactacc tcaagaacca
     3841 gcccatgtac gtgttcagaa agaccgagct caagcacagc aagacagagc ttaacttcaa
     3901 ggagtggcag aaggccttca ccgacgtgat gggcatggat gagctgtaca agtaactcga
     3961 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     4021 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4081 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4141 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4201 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4261 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4321 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4381 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4441 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4501 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4561 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4621 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4681 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4801 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4861 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4921 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4981 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5041 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5101 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5161 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5221 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5281 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5341 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5401 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5461 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5521 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5581 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5641 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5701 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5761 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5821 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5881 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5941 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6001 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6061 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6181 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6241 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6301 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6361 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6421 ga
//
