LOCUS       sequence-413187-        6235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of y-ATOM biosensor for SH2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  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 6235)
  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-MBP-yATOM_SH2
FEATURES             Location/Qualifiers
     source          1..6235
                     /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             642..1742
                     /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     1716..1739
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1755..1778
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     1886..1907
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2013..2034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3023..3144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3060..3079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3114..3133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3151..3606)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3288..3309)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3500..3519
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3633..3737
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3739..4096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3739..3930
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3762..3782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3947..4082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4131..4925
                     /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             4131..4925
                     /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(4185..4204)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4795..4814
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5113..5132)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5157..5204
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5533..6121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6022..6041
                     /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 catgaaaact gaagaaggta
      661 aactggtaat ctggattaac ggcgataaag gctataacgg tctcgctgaa gtcggtaaga
      721 aattcgagaa agataccgga attaaagtca ccgttgagca tccggataaa ctggaagaga
      781 aattcccaca ggttgcggca actggcgatg gccctgacat tatcttctgg gcacacgacc
      841 gctttggtgg ctacgctcaa tctggcctgt tggctgaaat caccccggac aaagcgttcc
      901 aggacaagct gtatccgttt acctgggatg ccgtacgtta caacggcaag ctgattgctt
      961 acccgatcgc tgttgaagcg ttatcgctga tttataacaa agatctgctg ccgaacccgc
     1021 caaaaacctg ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg
     1081 cgctgatgtt caacctgcaa gaaccgtact tcacctggcc gctgattgct gctgacgggg
     1141 gttatgcgtt caagtatgaa aacggcaagt acgacattaa agacgtgggc gtggataacg
     1201 ctggcgcgaa agcgggtctg accttcctgg ttgacctgat taaaaacaaa cacatgaatg
     1261 cagacaccga ttactccatc gcagaagctg cctttaataa aggcgaaaca gcgatgacca
     1321 tcaacggccc gtgggcatgg tccaacatcg acaccagcaa agtgaattat ggtgtaacgg
     1381 tactgccgac cttcaagggt caaccatcca aaccgttcgt tggcgtgctg agcgcaggta
     1441 ttaacgccgc cagtccgaac aaagagctgg caaaagagtt cctcgaaaac tatctgctga
     1501 ctgatgaagg tctggaagcg gttaataaag acaaaccgct gggtgccgta gcgctgaagt
     1561 cttacgagga agagttggcg aaagatccac gtattgccgc cactatggaa aacgcccaga
     1621 aaggtgaaat catgccgaac atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg
     1681 cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga
     1741 ctaattcgag ctcgctggaa gttctgttcc aggggcccaa ggtaccagga gctgcgacac
     1801 cggtaactgc agacatgctg cccgacaacc actacctgag ctatcagtcc gttctgagca
     1861 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1921 tcactctcgg catggacgag ctgtacaagg gcggtggatc cggtggtatg gtgagcaagg
     1981 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2041 gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc
     2101 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2161 tgggctacgg cctagcctgc ttcagccgct accccgacca catgaagcag cacgacttct
     2221 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg
     2281 gtacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2341 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2401 acttcaacag ccacaacgtc tatatcacgg ccgacaagtc tccggttcag gaattcactg
     2461 taccatattc ttcttctact gctaccatca gcggcctgag cccgggtgtc gactatacca
     2521 tcactgtata cgcatggggt gaagactctg ctggttacat gttcatgtac agcccaatct
     2581 cgattaacta ccgtaccgga ggttcaggca gtggtggtgc tagcggtggt gcgactggag
     2641 gcagtggcgg tggatcttct gttccgacca aactggaagt tgttgctgcg accccgacta
     2701 gcctgctgat cagctgggat gcgccgatgt cttcttcttc tgtttactat taccgtatca
     2761 cgtacggtga aacgggtggt aaccagaaga acggcatcaa ggctaacttc aagatccgcc
     2821 acaacgttga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2881 gcgacggccc cgtgctgtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     2941 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3001 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3061 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3121 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3181 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3241 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3301 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3361 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3421 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3481 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3541 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3601 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3661 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3721 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3781 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3841 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3901 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3961 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4021 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4081 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4141 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4201 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4261 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4321 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4381 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4441 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4501 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4561 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4621 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4681 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4741 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4801 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4861 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4921 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4981 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5041 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5101 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5161 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5221 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5281 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5341 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5401 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5461 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5521 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5581 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5641 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5701 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5761 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5821 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5881 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5941 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6001 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6061 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6121 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6181 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
