LOCUS       sequence-224299-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PSD-targeted AKARet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  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 7287)
  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: AKARet-PSD
FEATURES             Location/Qualifiers
     source          1..7287
                     /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             join(3659..3661,3662..3664,3665..4378)
                     /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(3659..3661,3662..3664,3665..4375)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4402..4523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4413..4432)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4467..4486
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4704..5159
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4791..4810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5001..5022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5226..5245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5345..5367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5405..5423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5491..5595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5596..5664,5665..6456)
                     /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(5814..5833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6627..7214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7115..7134
                     /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 gctcgagaag ttttctcaag aacagatcgg
     2881 cgaaaacatt gtgtgcaggg tcatttgtac cacgggtcaa attcccatcc gagatttgtc
     2941 agctgatatt tcacaagtgc ttaaggaaaa acgatccata aagaaagttt ggacatttgg
     3001 tagaaaccca gcctgtgact atcatttagg aaacatttca agactgtcaa ataagcattt
     3061 ccaaatacta ctaggagaag acggtaacct tttattgaat gacatttcca ctaatgggac
     3121 ctggttaaat gggcaaaaag tcgagaagaa cagcaatcag ttactgtccc aaggtgatga
     3181 aataaccgtt ggtgtaggcg tggaatcaga tattttatct ctggtcattt tcataaacga
     3241 caaatttaag cagtgcctgg agcagaacaa agttgatcgc ggtaccagtg ctggtggtag
     3301 tgctggtggt agtgctggtg gtagtgctgg tggttccggc agtgctggtg gtagtgctgg
     3361 tggtagtacc agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggtag
     3421 tgctggtggt tccggcagtg ctggtggtag tgctggtggt agtaccagtg ctggtggtag
     3481 tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggttccg gcagtgctgg
     3541 tggtagtgct ggtggtagta ccagtgctgg tggtagtgct ggtggtagtg ctggtggtag
     3601 tgctggtggt agtgctggtg gttccggatt gaggcgcgcg acgctggttg acacgcgtat
     3661 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3721 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3781 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3841 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3901 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3961 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4021 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4081 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4141 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4201 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4261 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4321 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagc
     4381 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     4441 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4501 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4561 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     4621 ataaggatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4681 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4861 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5041 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5101 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     5161 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     5221 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5281 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5341 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5401 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5461 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5521 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5581 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5641 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5701 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5761 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt gctgctatgt
     5821 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5881 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5941 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6001 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6061 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6121 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6181 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6241 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6301 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6361 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6421 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6481 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6541 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6661 ttgcaaacaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7201 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7261 cttttgctca catggctcga cagatct
//
