LOCUS       sequence-413185-        4344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SH2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4344)
  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 4344)
  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-SH2
FEATURES             Location/Qualifiers
     source          1..4344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        439..742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        743..946
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     897..917
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1510..1631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1547..1566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1601..1620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1638..2093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1775..1796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1987..2006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2120..2224
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2226..2583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2226..2417
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2249..2269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2434..2569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2618..3412
                     /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             2618..3412
                     /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(2672..2691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3282..3301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3600..3619)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3644..3691
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4020..4344,1..264)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
       61 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      301 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      361 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
      421 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      481 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      541 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      601 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      661 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      721 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      781 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      841 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      901 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      961 tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat tcggccacca
     1021 tgccaagcaa ctacatcacg ccagtcaaca gtctggagaa acactcctgg taccatgggc
     1081 ctgtgtcccg caatgccgct gagtatctgc tgagcagcgg gatcaatggc agcttcttgg
     1141 tgcgtgagag tgagagcagt cctggccaga ggtccatctc gctgagatac gaagggaggg
     1201 tgtaccatta caggatcaac actgcttctg atggcaagct ctacgtctcc tccgagagcc
     1261 gcttcaacac cctggccgag ttggttcatc atcattcaac ggtggccgac gggctcatca
     1321 ccacgctcca ttatccagcc ccaaagcgca acaagcccac tgtctatggt gtgtccccca
     1381 actactgagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     1441 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1501 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1621 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1681 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1741 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1801 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     1861 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     1921 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     1981 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2041 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2221 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2461 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2521 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2581 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2641 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2701 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2761 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     2821 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2881 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2941 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3001 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3061 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3121 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3181 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3241 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3301 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3361 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3421 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3481 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3541 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     3601 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3661 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3721 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3781 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     3841 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     3901 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4081 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4141 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4321 gcgcagcggt cgggctgaac gggg
//
