LOCUS       sequence-413182-        5176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of y-ATOM biosensor for SH2 localized to 
            endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5176)
  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 5176)
  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-PRLSS-yATOM_SH2-KDEL
FEATURES             Location/Qualifiers
     source          1..5176
                     /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"
     primer_bind     806..827
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(933..954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1964..2085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2001..2020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2055..2074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2092..2547)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2229..2250)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2441..2460
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2574..2678
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2680..3037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2680..2871
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2703..2723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2888..3023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3072..3866
                     /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             3072..3866
                     /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(3126..3145)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3736..3755
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4054..4073)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4098..4145
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4474..5062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4963..4982
                     /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 catgaacatc aaaggatcgc
      661 catggaaagg gtccctcctg ctgctgctgg tgtcaaacct gctcctgtgc cagagcgtgg
      721 caccggtaac tgcagacctg cccgacaacc actacctgag ctatcagtcc gttctgagca
      781 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      841 tcactctcgg catggacgag ctgtacaagg gcggtggatc cggtggtatg gtgagcaagg
      901 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      961 gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc
     1021 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1081 tgggctacgg cctagcctgc ttcagccgct accccgacca catgaagcag cacgacttct
     1141 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg
     1201 gtacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1261 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1321 acttcaacag ccacaacgtc tatatcacgg ccgacaagtc tccggttcag gaattcactg
     1381 taccatattc ttcttctact gctaccatca gcggcctgag cccgggtgtc gactatacca
     1441 tcactgtata cgcatggggt gaagactctg ctggttacat gttcatgtac agcccaatct
     1501 cgattaacta ccgtaccgga ggttcaggca gtggtggtgc tagcggtggt gcgactggag
     1561 gcagtggcgg tggatcttct gttccgacca aactggaagt tgttgctgcg accccgacta
     1621 gcctgctgat cagctgggat gcgccgatgt cttcttcttc tgtttactat taccgtatca
     1681 cgtacggtga aacgggtggt aaccagaaga acggcatcaa ggctaacttc aagatccgcc
     1741 acaacgttga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1801 gcgacggccc cgtgctgggt agcggtaaag acgagctgta aagcggccgc gactctagat
     1861 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1921 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1981 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2041 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2101 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2161 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2221 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2281 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2341 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2401 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2461 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2521 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2581 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2641 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2701 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2761 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2881 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2941 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3001 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3061 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3121 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3181 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3241 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3301 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3361 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3421 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3481 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3541 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3601 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3661 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3721 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3781 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3841 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3901 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3961 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4021 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4081 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4141 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4201 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4261 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4321 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4381 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4441 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4501 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4561 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4621 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4681 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4741 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4801 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4861 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4921 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4981 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5041 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5101 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5161 tattaccgcc atgcat
//
