LOCUS       sequence-413180-        4458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SH2 localized to endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  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 4458)
  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-SH2-KDEL
FEATURES             Location/Qualifiers
     source          1..4458
                     /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"
     polyA_signal    1246..1367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1283..1302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1337..1356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1374..1829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1511..1532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1723..1742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1856..1960
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1962..2319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1962..2153
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1985..2005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2170..2305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2354..3148
                     /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             2354..3148
                     /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(2408..2427)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3018..3037
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3336..3355)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3380..3427
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3756..4344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4245..4264
                     /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 caccggtctc aggtgctcca agcaactaca tcacgccagt caacagtctg gagaaacact
      781 cctggtacca tgggcctgtg tcccgcaatg ccgctgagta tctgctgagc agcgggatca
      841 atggcagctt cttggtgcgt gagagtgaga gcagtcctgg ccagaggtcc atctcgctga
      901 gatacgaagg gagggtgtac cattacagga tcaacactgc ttctgatggc aagctctacg
      961 tctcctccga gagccgcttc aacaccctgg ccgagttggt tcatcatcat tcaacggtgg
     1021 ccgacgggct catcaccacg ctccattatc cagccccaaa gcgcaacaag cccactgtct
     1081 atggtgtgtc ccccaactac ggtagcggta aagacgagct gtgagcggcc gcgactctag
     1141 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1201 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1321 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1381 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1441 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1501 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1561 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1621 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1681 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1741 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     1801 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     1861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1921 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     1981 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2041 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2221 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2281 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2341 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2401 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2461 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2521 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2581 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2641 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2701 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2761 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2821 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2881 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2941 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3001 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3061 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3121 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3181 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3241 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3301 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3361 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3421 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3481 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3541 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3901 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4381 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4441 cgtattaccg ccatgcat
//
