AAV-PCaMKII-Mt-mEGFP-CREB (S133A)

LOCUS       AAV-PCaMKII-Mt-mEGFP-CREB (S133A)               7038 bp DNA     circular SYN 02-APR-2026
DEFINITION  Forebrain principle neuron specific expression of GCaMP6f Ca sensor 
            and physically separate nuclear localized dTomato fluorophore .
ACCESSION   .
VERSION     .
KEYWORDS    2478-AAV-PCaMKII-Mt-mEGFP-CREB (S133A)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  TITLE     Ting GCaMP6 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7038)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 31, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        164..1452
                     /label=mouse aCaMKII promoter full length
     primer_bind     1372..1390
                     /label=AAV-aCaMKKII_F
     CDS             1480..1659
                     /codon_start=1
                     /label=Mito-advanced Sequence
                     /translation="MAQRLLLRRFLASVISRKPSQGQWPPLTSRALQTPQCSPGGLTVT
                     PNPARTIYTTRISLT"
     primer_bind     1647..1692
                     /label=mNeoGreen_F_AgeI
     CDS             1675..2391
                     /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(1720..1741)
                     /label=pEGFP-N-3'
     primer_bind     complement(1720..1741)
                     /label=EGFP-N
     primer_bind     1981..2000
                     /label=EXFP_F
     primer_bind     complement(1981..2000)
                     /label=EXFP-R
     primer_bind     complement(2274..2304)
                     /label=A206K_R
     primer_bind     2285..2316
                     /label=A206K_F
     primer_bind     2328..2349
                     /label=EGFP-C
     primer_bind     complement(2371..2415)
                     /label=mEGFP_R EcoRI
     primer_bind     2393..2415
                     /label=EcoRI CREB F
     CDS             2404..3426
                     /codon_start=1
                     /label=CREB
                     /translation="MTMDSGADNQQSGDAAVTEAESQQMTVQAQPQIATLAQVSMPAAH
                     ATSSAPTVTLVQLPNGQTVQVHGVIQAAQPSVIQSPQVQTVQSSCKDLKRLFSGTQIST
                     IAESEDSQESVDSVTDSQKRREILSRRPAYRKILNDLSSDAPGVPRIEEEKSEEETSAP
                     AITTVTVPTPIYQTSSGQYIAITQGGAIQLANNGTDGVQGLQTLTMTNAAATQPGTTIL
                     QYAQTTDGQQILVPSNQVVVQAASGDVQTYQIRTAPTSTIAPGVVMASSPALPTQPAEE
                     AARKREVRLMKNREAARECRRKKKEYVKCLENRVAVLENQNKTLIEELKALKDLYCHKS
                     D"
     primer_bind     complement(2785..2812)
                     /label=S133A_R2
     primer_bind     complement(2789..2812)
                     /label=WT_R
     primer_bind     2813..2843
                     /label=S142-3A_F
     primer_bind     2813..2843
                     /label=WT_F
     iDNA            2969..2989
                     /label=shCREB!
     primer_bind     complement(2997..3013)
                     /label=CREB_R
     iDNA            3392..3412
                     /label=shCREB?
     primer_bind     complement(3403..3453)
                     /label=A-CREB_R_SpeI
     primer_bind     complement(3437..3468)
                     /label=CREB_EcoRV_R
     misc_feature    3457..4045
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3510..3530)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3928..3939)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4078..4189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4301..4441
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4516..4971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5253..5357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5358..6218
                     /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(5576..5595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6389..6977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaat taacattatg gccttaggtc
      181 acttcatctc catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga
      241 gcttcctcag tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca
      301 gattactaat cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg
      361 aactcattag ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc
      421 tacaccaccc gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct
      481 ctccagtcct accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc
      541 aaatcttcat ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag
      601 gggccctgct gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca
      661 gcattctggg tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat
      721 ggctaaaaag cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca
      781 gaccaattcc cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga
      841 gagatgaatt agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca
      901 atacctgtct cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag
      961 gaccatccca gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac
     1021 ttaatcccat cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag
     1081 cgatgatcac ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc
     1141 accctggggg aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat
     1201 ataaggccag ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg
     1261 cagttaccgg ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg
     1321 ctgacgaagg ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc
     1381 gtcagtcaag ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt
     1441 tctgggggca gcgggggatc ctgatcccct agggccacca tggctcagcg acttcttctg
     1501 aggaggttcc tggcctctgt catctccagg aagccctctc agggtcagtg gccacccctc
     1561 acttccagag ccctccagac cccacaatgc agtcctggtg gcctgactgt aacacccaac
     1621 ccagcccgga caatatacac cacgaggatc tccttgacac ctaggccacc ggttatggtg
     1681 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1741 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1801 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1861 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1921 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1981 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2041 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2101 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2161 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2221 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2281 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2341 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctctacaa ggaattcgct
     2401 agcatgacca tggactctgg agcagacaac cagcagagtg gagatgctgc tgtaacagaa
     2461 gctgaaagtc aacaaatgac agttcaagcc cagccacaga ttgccacatt agcccaggta
     2521 tccatgccag cagctcatgc gacgtcatct gctcccactg taaccttagt gcagctgccc
     2581 aatgggcaga cagtccaggt ccatggggtc atccaggcgg cccagccatc agttattcag
     2641 tctccacaag tccaaacagt tcagtcttcc tgtaaggact taaaaagact tttctccgga
     2701 actcagattt caactattgc agaaagtgaa gattcacagg agtctgtgga tagtgtaact
     2761 gattcccaaa aacgaaggga aatcctttca aggaggcctg catacaggaa aattttgaat
     2821 gacttatctt ctgatgcacc aggggtgcca aggattgaag aagaaaaatc agaagaagag
     2881 acttcagccc ctgccatcac cactgtaaca gtgccaaccc cgatttacca aactagcagt
     2941 gggcagtata ttgccattac ccagggagga gcaatacagc tggctaacaa tggtaccgat
     3001 ggggtacagg gcctgcagac attaaccatg accaatgcag ctgccactca gccgggtact
     3061 accattctac aatatgcaca gaccactgat ggacagcaga ttctagtgcc cagcaaccaa
     3121 gttgttgttc aagctgcctc tggtgatgta caaacatacc agattcgcac agcacccact
     3181 agcaccattg cccctggagt tgttatggcg tcctccccag cacttcctac acagcctgct
     3241 gaagaagcag cacgaaagag agaggttcgt ctaatgaaga acagggaagc agcaagagaa
     3301 tgtcgtagaa agaagaaaga atatgtgaaa tgtttagaga acagagtggc agtgcttgaa
     3361 aaccaaaaca aaacattgat tgaggagcta aaagcactta aggaccttta ctgccacaag
     3421 tcagattaaa ctagttgaga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3481 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3541 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3601 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3661 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3721 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3781 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3841 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3901 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3961 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4021 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctcgactg
     4081 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4141 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4201 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4261 aagacaatag caggcatgca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4321 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4381 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4441 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4501 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     4861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4921 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4981 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5041 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5101 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5161 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5221 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5281 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5341 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5461 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5521 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5581 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5641 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5761 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5881 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5941 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6181 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7021 ggccttttgc tcacatgt
//