LOCUS       sequence-413186-        6262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of c-ATOM biosensor for HRas with nuclear 
            export signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6262)
  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 6262)
  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-NES-MBP-cATOM(Ras)
FEATURES             Location/Qualifiers
     source          1..6262
                     /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             675..1775
                     /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="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1749..1772
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1788..1811
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     1919..1940
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2046..2067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3050..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3178..3633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3315..3336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3527..3546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3660..3764
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3766..4123
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3766..3957
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3789..3809)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3974..4109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4158..4952
                     /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             4158..4952
                     /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(4212..4231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4822..4841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5140..5159)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5184..5231
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5560..6148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6049..6068
                     /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 gctagcgcca
      601 ccatgaacct ggtggacctc caaaagaagc tggaggagct ggagctggac gagcagcagg
      661 gaccggtcgc caccatgaaa actgaagaag gtaaactggt aatctggatt aacggcgata
      721 aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag
      781 tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg
      841 atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc
      901 tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg
      961 atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc
     1021 tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc
     1081 tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt
     1141 acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca
     1201 agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc
     1261 tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag
     1321 ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca
     1381 tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat
     1441 ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc
     1501 tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata
     1561 aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc
     1621 cacgtattgc cgccactatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc
     1681 agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc
     1741 agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgctg gaagttctgt
     1801 tccaggggcc caaggtacca ggagctgcga caccggtaac tgcagacatg ctgcccgaca
     1861 accactacct gagcactcag tccgttctga gcaaagaccc caacgagaag cgcgatcaca
     1921 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1981 agggcggtgg atccggtggt atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2041 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg
     2101 gcgagggcga tgccaccaac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     2161 tgcccgtgcc ctggcccacc ctcgtgacca ccctcagctg gggcgtgcag tgcttcgcgc
     2221 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2281 tccaggagcg caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga
     2341 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2401 acggcaacat cctggggcac aagctggagt acaactatat tagcgataac gtctatatca
     2461 cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa gatccgccac aacgttgagg
     2521 acggtgtgga ctacacgatt accgtgtatg cgtggggttg gcatggccag gtttattact
     2581 atatgggtag cccaatctcg attaactacc gtaccggagg ttcaggcagt ggtggtgcta
     2641 gcggtggtgc gactggaggc agtggcggtg gatcttctgt tccgaccaaa ctggaggtgg
     2701 ttgccgcaac tcccacgagt ctgctgatct catgggatgc tccagccgtt accgtcgatt
     2761 attacgtcat cacctatggc gaaacaggcg gcaactctcc ggtacagaag tttgaagtgc
     2821 ctgggtccaa aagcactgcg accatttcgg gtttaaaacc gggcagcgtg cagctcgccg
     2881 accactacca gcagaacacc cccatcggcg acggccccgt gctgtaaagc ggccgcgact
     2941 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3001 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3181 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3241 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3301 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3361 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3421 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3481 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3541 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3601 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3661 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3721 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3781 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3841 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3901 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3961 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4021 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4081 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4141 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4201 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4261 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4321 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4381 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4441 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4501 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4561 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4621 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4681 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4741 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4801 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4861 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4921 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4981 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5041 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5101 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5161 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5221 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5281 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5341 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5401 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5461 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5521 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5581 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5641 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5701 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5761 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5821 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5881 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5941 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6001 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6061 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6181 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6241 taaccgtatt accgccatgc at
//
