LOCUS       sequence-413171-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of r-ATOM biosensor for WDR5 with nuclear 
            localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 6238)
  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: "Nat 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-NLS-MBP-rATOM(WDR5)
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             645..665
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             681..1778
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1752..1775
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1791..1814
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      1842..1851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3026..3147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3063..3082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3117..3136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3154..3609)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3291..3312)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3503..3522
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3636..3740
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3742..4099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3742..3933
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3765..3785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3950..4085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4134..4928
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4134..4928
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4188..4207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4798..4817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5116..5135)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5160..5207
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5536..6124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6025..6044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgccac catgccgaag aagaaacgca
      661 aagtgggagg ttcaggcagt aaaactgaag aaggtaaact ggtaatctgg attaacggcg
      721 ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta
      781 aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt gcggcaactg
      841 gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg
      901 gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat ccgtttacct
      961 gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat
     1021 cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa gagatcccgg
     1081 cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac
     1141 cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgaaaacg
     1201 gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct
     1261 tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac tccatcgcag
     1321 aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca
     1381 acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc aagggtcaac
     1441 catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag
     1501 agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg gaagcggtta
     1561 ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag
     1621 atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc
     1681 cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc
     1741 gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg ctggaagttc
     1801 tgttccaggg gcccaaggta ccaggagctg cgacaccggt cgccaccatg gtgtctaagg
     1861 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     1921 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     1981 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     2041 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gatttcttta
     2101 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2161 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     2221 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     2281 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg
     2341 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     2401 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     2461 gaatcaagga ggccgacggc aactctccgg tacagaagtt taaagtgcct gggtccaaaa
     2521 gcactgcgac catttcgggc ttaaaaccgg gtgtggacta cacgattacc gtgtatgcgt
     2581 atcaaggtgg tggacgctgg catccgtatg ggtactatag cccgatctcg attaactacc
     2641 gtaccggagg ttcaggcagt ggtggtgcta gcggtggtgc gactggaggc agtggcggtg
     2701 gatcttctgt tccgaccaaa ttggaggtgg ttgccgcaac tcccacgagt ctgctgatct
     2761 catgggatgc tccagccgtt accgtcgtac actacgtcat cacctatggc gaaacaggca
     2821 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     2881 aactggggca caaacttaat taaagcggcc gcgactctag atcataatca gccataccac
     2941 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3001 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3061 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3121 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3181 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3241 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3301 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3361 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3421 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3481 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3541 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3601 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3661 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3721 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3781 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3901 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3961 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4021 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4081 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4441 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4921 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4981 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5041 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5101 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5161 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5221 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5281 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5341 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5401 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5461 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5521 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5581 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5641 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5701 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5761 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5821 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5881 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5941 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6001 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6061 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6121 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6181 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
