LOCUS       sequence-272089-        5458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone, mEGFP-CREB donor for G-CREB with S133A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5458)
  AUTHORS   Laviv T, Scholl B, Parra-Bueno P, Foote B, Zhang C, Yan L, Hayano Y,
            Chu J, Yasuda R
  TITLE     In Vivo Imaging of the Coupling between Neuronal and CREB Activity 
            in the Mouse Brain.
  JOURNAL   Neuron. 2019 Dec 17. pii: S0896-6273(19)31037-2. doi: 
            10.1016/j.neuron.2019.11.028.
  PUBMED    31883788
REFERENCE   2  (bases 1 to 5458)
  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: "Neuron. 
            2019 Dec 17. pii: S0896-6273(19)31037-2. doi: 
            10.1016/j.neuron.2019.11.028."
COMMENT     Sequence Label: AAV-SYN_mEGFP-CREB (S133A)
FEATURES             Location/Qualifiers
     source          1..5458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        210..657
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(688..690,691..693,694..1404)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(733..754)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(994..1013)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1341..1362
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   2721..2861
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2721..2850
                     /label=AAV2 ITR
     rep_origin      2936..3391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3023..3042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3233..3254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3408..3427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3527..3549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3587..3605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3673..3777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3778..3846,3847..4638)
                     /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(3996..4015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4809..5397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5298..5317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtattacc gccatgcatt agttattaat agtaatcaat
      181 tacggggtct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
      241 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
      301 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
      361 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
      421 agccacagcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
      481 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
      541 gcgaggcgcg agataggggg gcgcggacgc gaccatctgc gctgcggcgc cggcgactca
      601 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
      661 aattggctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      721 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      781 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      841 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      901 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      961 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1021 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1081 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1141 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1201 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1261 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     1321 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1381 ctcggcatgg acgagctgta caagcctacc cctaccccta ccatgaccat ggactctgga
     1441 gcagacaacc agcagagtgg agatgctgct gtaacagaag ctgaaagtca acaaatgaca
     1501 gttcaagccc agccacagat tgccacatta gcccaggtat ccatgccagc agctcatgcg
     1561 acgtcatctg ctcccactgt aaccttagtg cagctgccca atgggcagac agtccaggtc
     1621 catggggtca tccaggcggc ccagccatca gttattcagt ctccacaagt ccaaacagtt
     1681 cagtcttcct gtaaggactt aaaaagactt ttctccggaa ctcagatttc aactattgca
     1741 gaaagtgaag attcacagga gtctgtggat agtgtaactg attcccaaaa acgaagggaa
     1801 atcctttcaa ggaggcctgc ctacaggaaa attttgaatg acttatcttc tgatgcacca
     1861 ggggtgccaa ggattgaaga agaaaaatca gaagaagaga cttcagcccc tgccatcacc
     1921 actgtaacag tgccaacccc gatttaccaa actagcagtg ggcagtatat tgccattacc
     1981 cagggaggag caatacagct ggctaacaat ggtaccgatg gggtacaggg cctgcagaca
     2041 ttaaccatga ccaatgcagc tgccactcag ccgggtacta ccattctaca atatgcacag
     2101 accactgatg gacagcagat tctagtgccc agcaaccaag ttgttgttca agctgcctct
     2161 ggtgatgtac aaacatacca gattcgcaca gcacccacta gcaccattgc ccctggagtt
     2221 gttatggcgt cctccccagc acttcctaca cagcctgctg aagaagcagc acgaaagaga
     2281 gaggttcgtc taatgaagaa cagggaagca gcaagagaat gtcgtagaaa gaagaaagaa
     2341 tatgtgaaat gtttagagaa cagagtggca gtgcttgaaa accaaaacaa aacattgatt
     2401 gaggagctaa aagcacttaa ggacctttac tgccacaagt cagattaaaa gcttgtggca
     2461 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     2521 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2581 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga cccaaattgc
     2641 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     2701 cgtgtggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2761 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2821 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2881 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3001 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3121 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3241 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3301 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3361 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3421 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3481 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3541 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3601 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3721 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3781 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3841 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3901 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3961 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4021 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4081 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4141 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4201 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4261 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4321 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4381 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4621 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4681 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4741 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4801 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4861 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4921 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4981 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5041 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5101 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5161 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5221 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5281 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5341 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5401 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
