LOCUS       sequence-224301-        7068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PSD-targeted EKARet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7068)
  AUTHORS   Tang S, Yasuda R
  TITLE     Imaging ERK and PKA Activation in Single Dendritic Spines during 
            Structural Plasticity.
  JOURNAL   Neuron. 2017 Mar 22;93(6):1315-1324.e3. doi: 
            10.1016/j.neuron.2017.02.032. Epub 2017 Mar 9.
  PUBMED    28285819
REFERENCE   2  (bases 1 to 7068)
  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.neuron.2017.02.032"; journalName: "Neuron"; date: 
            "2017-03-22- 22"; volume: "93"; issue: "6"; pages: "1315-1324.e3"
COMMENT     Sequence Label: EKARet-PSD
FEATURES             Location/Qualifiers
     source          1..7068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1034..1053
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1058..1067
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2111..2120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2135..2137,2138..2140,2141..2851)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITLGMDELYK"
     CDS             3026..3373
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             join(3440..3442,3443..3445,3446..4159)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3440..3442,3443..3445,3446..4156)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4183..4304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4248..4267
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4485..4940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4572..4591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4782..4803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5007..5026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5126..5148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5186..5204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5272..5376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5377..5445,5446..6237)
                     /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     complement(5595..5614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6408..6995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6896..6915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggcg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata gggtggcgcc accatggact gtctctgtat
     1081 agtgacaacc aagaaatacc gctaccaaga tgaagacacg ccccctctgg aacacagccc
     1141 ggcccacctc cccaaccagg ccaattctcc ccctgtgatt gtcaacacgg acaccctaga
     1201 agccccagga tatgagttgc aggtgaatgg aacagagggg gagatggagt atgaggagat
     1261 cacattggaa aggggtaact caggtctggg cttcagcatc gcaggtggca ctgacaaccc
     1321 gcacatcggt gacgacccgt ccatttttat caccaagatc attcctggtg gggctgcagc
     1381 ccaggatggc cgcctcaggg tcaatgacag catcctgttt gtaaatgaag tggatgttcg
     1441 ggaggtgacc cattcagctg cggtggaggc cctcaaagag gcaggttcca tcgttcgcct
     1501 ctatgtcatg cgccggaaac ccccagccga aaaggtcatg gagatcaaac tcatcaaagg
     1561 gcctaaagga cttggcttca gcattgcggg gggcgttggg aaccagcaca tccctggaga
     1621 taacagcatc tatgtaacaa agatcatcga aggaggtgct gcccacaagg atggcaggtt
     1681 gcagattgga gacaagatcc tggcggtcaa cagtgtgggg ctggaggacg tcatgcacga
     1741 ggatgccgtg gcagccctga agaacacata tgacgttgtg tacctaaagg tggccaagcc
     1801 cagcaatgcc tacctgagtg acagctatgc tcccccagac atcacaacct cgtattctca
     1861 gcacctggac aatgagatca gtcatagcag ctacttgggc actgactacc ccacagccat
     1921 gacccccact tcccctcggc gctactcccc tgtggccaag gacctgctgg gggaggaaga
     1981 cattccccgg atctatcaca aagtgaaacg tgtcattgaa gacctctcag gcccctacat
     2041 ctgggtccca gcccgagaga gactcggaat tctgcagtcg acggtaccgc gggcccggga
     2101 tccaccggtc gccaccatgg catgccaggc tagcatggtg agcaagggcg aggagctgtt
     2161 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2221 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctcatctg
     2281 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     2341 gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2401 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2461 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2521 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact ggaacagcgt
     2581 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2641 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2701 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgtt
     2761 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcctga ccgccgccgg
     2821 gatcactctc ggcatggacg agctgtacaa gctcgagatg gcggacgagg agaagctgcc
     2881 gcccggctgg gagaagcgca tgagccgcag ctcaggccga gtgtactact tcaaccacat
     2941 cactaacgcc agccagtggg agcggcccag cggcaacagc agcagtggtg gcaaaaacgg
     3001 gcagggggag cctgccaggg gtaccagtgc tggtggtagt gctggtggta gtgctggtgg
     3061 tagtgctggt ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac
     3121 cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg
     3181 ttccggcagt gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg
     3241 tagtgctggt ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc
     3301 tggtggtagt accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg
     3361 tagtgctggt ggttccggac cagatgtccc tagaactcca gtggataaag caaagctgtc
     3421 attccaattt ccgacgcgta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3481 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3541 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3601 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3661 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3721 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3781 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3841 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3901 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3961 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4021 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4081 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4141 ggacgagctg tacaagtaag cggccgcttc gagcagacat gataagatac attgatgagt
     4201 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4261 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4321 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     4381 tctacaaatg tggtaaaatc gataaggatc cgggctggcg taatagcgaa gaggcccgca
     4441 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4801 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4921 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4981 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5041 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5101 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5161 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5221 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5281 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5581 acttttaaag tgctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6421 tttctgcgcg taatctgctg cttgcaaaca aaaaaccacc gctaccagcg gtggtttgtt
     6481 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6541 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6601 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6661 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6721 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6781 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6841 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6901 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6961 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7021 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
