LOCUS       sequence-401050-        6402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for camKII promoter (1.3kb)-driven smFP_FLAG (FLAG-tagged
            spaghetti monster) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6402)
  AUTHORS   Watakabe A, Tani T, Abe H, Skibbe H, Ichinohe N, Yamamori T
  TITLE     Serial Two-Photon Tomography of the Whole Marmoset Brain for 
            Neuroanatomical Analyses.
  JOURNAL   J Vis Exp. 2025 Jan 17;(215). doi: 10.3791/67694.
  PUBMED    39895625
REFERENCE   2  (bases 1 to 6402)
  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: "J Vis 
            Exp."; date: "2025-01-17"; volume: "(215). doi"; pages: " 
            10.3791/67694"
COMMENT     Sequence Label: pAAVCam1.3_smFP_FLAG
FEATURES             Location/Qualifiers
     source          1..6402
                     /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        145..1433
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1457..1466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1466..1489
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1493..1516
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1520..1543
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2072..2095
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2102..2125
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2138..2161
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2168..2191
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2402..2425
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2429..2452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2456..2479
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2529..3117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2582..2602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3000..3011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3149..3625
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3348..3367)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3665..3805
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3665..3794
                     /label=AAV2 ITR
     rep_origin      3880..4335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4352..4371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4471..4493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4531..4549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4617..4721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4722..4790,4791..5582)
                     /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(4940..4959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5753..6341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6242..6261
                     /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 gcgtcattat ggccttaggt cacttcatct ccatggggtt
      181 cttcttctga ttttctagaa aatgagatgg gggtgcagag agcttcctca gtgacctgcc
      241 cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa tcttaaattc
      301 catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta gggcagatga
      361 ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc cgcccagccc
      421 tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc taccttgacg
      481 ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca tggtccctct
      541 gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg
      601 tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg gtctgtttgg
      661 tacttcttct cacgataagg tggcggtgtg atatgcacaa tggctaaaaa gcagggagag
      721 ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc ccagaggaag
      781 caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat tagcttcccc
      841 tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc tcttcagagg
      901 agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc agaatgtggc
      961 acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca tcccgtctgt
     1021 taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca cttgtggact
     1081 aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg gaacagggtc
     1141 cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca gggcaaagag
     1201 gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg gggcaacggg
     1261 aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag gctcgcgagg
     1321 ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa gccggttctc
     1381 cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc agcatcgatt
     1441 gaattcaatt gcggccgcca ccatggacta caaggacgac gacgataaag gagattacaa
     1501 agatgacgat gataagggcg attataagga cgacgatgac aagggcggcg tgtcgaaagg
     1561 cgaggagctg ttcacaggcg tggtccccat cctggttgag ctggacggtg acgttaacgg
     1621 ccacaaattt agcgtacgtg gtgagggcga gggtgacgct accaacggaa agctgaccct
     1681 gaaattcatc tgcacaactg gcaagctgcc ggtgccctgg cccacgctgg taaccactct
     1741 gggcggaggc gttcaatgtt tcagtcgtta cccggatcac atgaagcagc atgacttctt
     1801 caagtcggcc atgcccgaag gctacgtgca ggagcgcacc atcagtttta aggatgacgg
     1861 cacctacaag actcgcgccg aggtcaagtt cgaaggagac accctggtta atcgtatcga
     1921 gctcaaaggc atcgatttca aggaggacgg aaatattctg ggccacaagc tggagtacaa
     1981 ttttaacagc cacaatgttt acattactgc cgacaagcag aagaacggca tcaaggctaa
     2041 tttcaagatt cgtcacaatg tggagggagg cgactataag gatgacgatg acaaacaaca
     2101 agactataag gacgatgatg acaaaggtca gcaaggcgat tacaaggacg atgacgataa
     2161 gcagcaagat tacaaggatg atgatgataa gggaggcgac ggaagcgtgc agctggcaga
     2221 tcattaccaa cagaacacgc ccattggtga cggtccagtc ctgctgcctg ataaccacta
     2281 cttgtccacc cagtcggtgc tgtcgaagga cccgaacgag aagcgtgacc acatggtgct
     2341 gctggagttc gtgaccgccg caggaatcac cctcggcatg gatgagcttt acaagggcgg
     2401 agattataaa gatgacgacg acaagggcga ctacaaagat gatgatgaca agggtgacta
     2461 caaagacgac gatgataaat aaaccggtgc tcgaggtacg gatccaattc gatatcaagc
     2521 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2941 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3121 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3181 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3241 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3301 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3361 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3421 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     3481 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3541 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3601 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3661 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3721 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3781 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3841 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4261 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4321 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4381 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4441 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4501 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4561 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5881 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
