LOCUS       sequence-345379-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of G-alpha-q fused to mNeonGreen-eDHFR(69K6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  AUTHORS   Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, 
            Atsuta-Tsunoda K, Aoki K, Tsukiji S
  TITLE     A chemogenetic platform for controlling plasma membrane signaling 
            and synthetic signal oscillation.
  JOURNAL   Cell Chem Biol. 2022 Sep 15;29(9):1446-1464.e10. doi: 
            10.1016/j.chembiol.2022.06.005. Epub 2022 Jul 13.
  PUBMED    35835118
REFERENCE   2  (bases 1 to 7357)
  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"; doi: 
            "10.1016/j.chembiol.2022.06.005"; journalName: "Cell Chem Biol"; 
            date: "2022-09-15- 15"; volume: "29"; issue: "9"; pages: 
            "1446-1464.e10"
COMMENT     Sequence Label: pCAGGS-mNG-eDHFR(69K6)-G-alpha-q
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..1126
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1050..1072
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1134..1153
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1187..1894
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3720..3740)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3720..3738)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3759..3775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3877..3896)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3942..3997
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3996..4015)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4358..4374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4382..4398
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4406..4412,4413..4430,4431..4436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4451..4472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4531..4726
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4577..4712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4732..4866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4769..4788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4823..4842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4934..4951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5105..5693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5185..5204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5864..6655,6656..6724))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6487..6506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6725..6829)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6860..7239
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      301 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
      361 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
      421 cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
      481 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
      541 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
      601 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
      661 cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg
      721 ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
      781 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
      841 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
      901 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
      961 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1021 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1081 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1141 gctggttatt gtgctgtctc atcattttgg caaagaattg gccaccatgg tgagcaaggg
     1201 cgaggaggat aacatggcct ctctcccagc gacacatgag ttacacatct ttggctccat
     1261 caacggtgtg gactttgaca tggtgggtca gggcaccggc aatccaaatg atggttatga
     1321 ggagttaaac ctgaagtcca ccaagggtga cctccagttc tccccctgga ttctggtccc
     1381 tcatatcggg tatggcttcc atcagtacct gccctaccct gacgggatgt cgcctttcca
     1441 ggccgccatg gtagatggct ccggctacca agtccatcgc acaatgcagt ttgaagatgg
     1501 tgcctccctt actgttaact accgctacac ctacgaggga agccacatca aaggagaggc
     1561 ccaggtgaag gggactggtt tccctgctga cggtcctgtg atgaccaact cgctgaccgc
     1621 tgcggactgg tgcaggtcga agaagactta ccccaacgac aaaaccatca tcagtacctt
     1681 taagtggagt tacaccactg gaaatggcaa gcgctaccgg agcactgcgc ggaccaccta
     1741 cacctttgcc aagccaatgg cggctaacta tctgaagaac cagccgatgt acgtgttccg
     1801 taagacggag ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcctt
     1861 taccgatgtg atgggcatgg acgagctcta caagtctggg tccgcagcca gtagcgccgg
     1921 aagtatgatc agtctgattg cggcgttagc ggtagatcgc gttatcggca tggaaaacgc
     1981 catgccgtgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct taaataaacc
     2041 cgtgattatg ggccgccata cctgggaatc aatcggtcgt ccgttgccag gacgcaaaaa
     2101 tattatcctc agcagtcaac cgggtacgga caaaaaaaag aaaaagaaag atcgcgtaac
     2161 gtgggtgaag tcggtggatg aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt
     2221 gattggcggc ggtcgcgttt atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac
     2281 gcatatcgac gcagaagtgg aaggcgacac ccatttcccg gattacgagc cggatgactg
     2341 ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag aactctcaca gctattgctt
     2401 tgagattctg gagcggcggc tgtactctgg tggcggaggc tcgggcggag gtgggtcggg
     2461 tggcggcgga tcttcagctg gagcttcgtt gtacaaggca agaggagccg ctgcgggtgc
     2521 cggaggcgct ggtaggagcg gcaaaaaggg gaagaaaggg aaaaagggca ccacaatgac
     2581 tctggagtcc atcatggcat cctccctgag cgaggaggcc aaggaagccc ggaggatcaa
     2641 cgacgagatc gagcggcagc tgcgcaggga caagcgcgac gcccgccggg agctcaagct
     2701 gctgctgctg gggacagggg agagtggcaa gtcgaccttc atcaagcaga tgaggatcat
     2761 ccacgggtcg ggctactctg acgaagacaa gcgcggcttc accaagctgg tgtatcagaa
     2821 catcttcacg gccatgcagg ccatgatcag agcgatggac acactcaaga tcccatacaa
     2881 gtatgaacac aataaggctc atgcacaatt ggttcgagag gttgatgtgg agaaggtgtc
     2941 tgcttttgag aatccatatg tagatgcaat aaagagcttg tggaatgatc ctggaatcca
     3001 ggagtgctac gacagacgac gggaatatca gttatctgac tctaccaaat actatctgaa
     3061 tgacttggac cgtgtagccg acccttccta tctgcctaca caacaagacg tgcttagagt
     3121 tcgagtcccc actacaggga tcatcgaata cccctttgac ttacaaagtg tcattttcag
     3181 aatggtcgat gtagggggcc tgaggtcaga gagaagaaaa tggatacact gctttgaaaa
     3241 tgtcacctcc atcatgtttc tagtagcgct tagcgaatat gatcaagttc ttgtggagtc
     3301 agacaatgag aaccgcatgg aggagagcaa agcactcttt agaacaatta tcacctaccc
     3361 ctggttccag aactcctctg tgattctgtt cttaaacaag aaagatcttc tagaggagaa
     3421 aatcatgtat tcccacctag tcgactactt cccagaatat gatggacccc agagagatgc
     3481 ccaggcagct cgagaattca tcctgaaaat gttcgtggac ctgaaccccg acagtgacaa
     3541 aatcatctac tcccacttca cgtgcgccac agataccgag aacatccgct tcgtctttgc
     3601 agccgtcaag gacaccatcc tgcagctgaa cctgaaggag tacaatctgg tctaacagtt
     3661 gctcgagagt acccgggata ctcgcggccg ccaccgcggt ggagctccag cttttgttcc
     3721 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     3781 aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3841 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3901 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3961 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4021 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     4081 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4141 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     4201 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     4261 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     4321 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4501 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     4561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4861 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5941 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6001 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6061 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6241 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6481 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6841 gaaaagtgcc acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg
     6901 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     6961 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     7021 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     7081 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     7141 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     7201 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
     7261 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     7321 taattatttt gtgcagcgat gggggcgggg ggggggg
//
