LOCUS       sequence-360827-        6821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to express nuclear localized E2-Crimson from CaMKIIa 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6821)
  AUTHORS   de Solis CA, Hosek MP, Holehonnur R, Ho A, Banerjee A, Luong JA, 
            Jones LE, Chaturvedi D, Ploski JE
  TITLE     Adeno-associated viral serotypes differentially transduce inhibitory
            neurons within the rat amygdala.
  JOURNAL   Brain Res. 2017 Oct 1;1672:148-162. doi: 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29.
  PUBMED    28764932
REFERENCE   2  (bases 1 to 6821)
  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: "Brain 
            Res."; date: "2017-10-1"; volume: "1672:148-162. doi"; pages: " 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29"
COMMENT     Sequence Label: pAAV-alphaCaMKII-E2-Crimson-3XNLS-pA
FEATURES             Location/Qualifiers
     source          1..6821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        228..1516
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     intron          1596..2071
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      2168..2177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2174..2848
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     2572..2591
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2861..2881
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2885..2905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2909..2929
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2971..2989)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2972..2989)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3009..3026)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3015..3126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3270..3399
                     /label=AAV2 ITR
     rep_origin      3770..4283
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3857..3876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4067..4088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4765..4784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4884..4906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4944..4962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5030..5134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5135..5203,5204..5995)
                     /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(5353..5372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6166..6754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6655..6674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
       61 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgagcgagcg cgcagagagg
      121 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      181 tatctacgta gccatgctct agaggatcct tcgaaaagct tgtataacat tatggcctta
      241 ggtcacttca tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca
      301 gagagcttcc tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa
      361 ctcagattac taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg
      421 agtgaactca ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta
      481 catctacacc acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc
      541 ctctctccag tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga
      601 gaccaaatct tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca
      661 ggaggggccc tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc
      721 gtcagcattc tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca
      781 caatggctaa aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca
      841 accagaccaa ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga
      901 aggagagatg aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg
      961 ggcaatacct gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct
     1021 caaggaccat cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc
     1081 ccacttaatc ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga
     1141 agagcgatga tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta
     1201 catcaccctg ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt
     1261 gtatataagg ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg
     1321 gaggcagtta ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg
     1381 gatgctgacg aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa
     1441 gctcgtcagt caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc
     1501 tagttctggg ggcagcgggg gatcccctac ggccggccac gcgttcgcga ccgcgggcta
     1561 gcggcgcgcc gagctcggat cctgagaact tcagggtgag tctatgggac ccttgatgtt
     1621 ttctttcccc ttcttttcta tggttaagtt catgtcatag gaaggggaga agtaacaggg
     1681 tacacatatt gaccaaatca gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc
     1741 ttttaatata cttttttgtt tatcttattt ctaatacttt ccctaatctc tttctttcag
     1801 ggcaataatg atacaatgta tcatgcctct ttgcaccatt ctaaagaata acagtgataa
     1861 tttctgggtt aaggcaatag caatatttct gcatataaat atttctgcat ataaattgta
     1921 actgatgtaa gaggtttcat attgctaata gcagctacaa tccagctacc attctgcttt
     1981 tattttatgg ttgggataag gctggattat tctgagtcca agctaggccc ttttgctaat
     2041 catgttcata cctcttatct tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct
     2101 ggcccatcac tttggcaaag cacgtgaatt cgaagatctg gccgcctcgg cctctagaac
     2161 tagtgccgcc accatggata gcactgagaa cgtcatcaag cccttcatgc gcttcaaggt
     2221 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgtgg gcgagggcaa
     2281 gccctacgag ggcacccaga ccgccaagct gcaagtgacc aagggcggcc ccctgccctt
     2341 cgcctgggac atcctgtccc cccagttctt ctacggctcc aaggcgtaca tcaagcaccc
     2401 cgccgacatc cccgactacc tcaagcagtc cttccccgag ggcttcaagt gggagcgcgt
     2461 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     2521 caccctcatc taccacgtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat
     2581 gcagaagaag actctgggct gggagccctc cactgagcgc aactaccccc gcgacggcgt
     2641 gctgaagggc gagaaccaca tggcgctgaa gctgaagggc ggcggccact acctgtgtga
     2701 gttcaagtcc atctacatgg ccaagaagcc cgtgaagctg cccggctacc actacgtgga
     2761 ctacaagctc gacatcacct cccacaacga ggactacacc gtggtggagc agtacgagcg
     2821 cgccgaggcc cgccaccacc tgttccagtc tcgagctgat ccaaaaaaga agagaaaggt
     2881 agatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag gatccaccgg
     2941 atctagataa cgtacgggaa acggccctat tctatagtgt cacctaaatg ctagagctcg
     3001 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3061 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3121 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3181 caagggggag gattgggaag acaatagcgc ggccgctcta gagcatggct acgtagataa
     3241 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3301 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3361 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc
     3421 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gattccgttg
     3481 caatggctgg cggtaatatt gttctggata ttaccagcaa ggccgatagt ttgagttctt
     3541 ctactcaggc aagtgatgtt attactaatc aaagaagtat tgcgacaacg gttaatttgc
     3601 gtgatggaca gactctttta ctcggtggcc tcactgatta taaaaacact tctcaggatt
     3661 ctggcgtacc gttcctgtct aaaatccctt taatcggcct cctgtttagc tcccgctctg
     3721 attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc aaccatagta cgcgccctgt
     3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4081 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4141 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4201 aacaaaatat taacgcttac aatttaaata tttgcttata caatcttcct gtttttgggg
     4261 cttttctgat tatcaaccgg ggtacatatg attgacatgc tagttttacg attaccgttc
     4321 atcgattctc ttgtttgctc cagactctca ggcaatgacc tgatagcctt tgtagagacc
     4381 tctcaaaaat agctaccctc tccggcatga atttatcagc tagaacggtt gaatatcata
     4441 ttgatggtga tttgactgtc tccggccttt ctcacccgtt tgaatcttta cctacacatt
     4501 actcaggcat tgcatttaaa atatatgagg gttctaaaaa tttttatcct tgcgttgaaa
     4561 taaaggcttc tcccgcaaaa gtattacagg gtcataatgt ttttggtaca accgatttag
     4621 ctttatgctc tgaggcttta ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt
     4681 tattggatgt tggaatcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4741 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4801 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4861 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4921 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4981 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5041 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5101 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5161 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5221 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5281 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5341 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5401 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5461 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5521 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5581 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5641 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5701 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5821 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6001 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6301 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6781 gttcctggcc ttttgctggc cttttgctca catgttcttt c
//
