LOCUS       SK0685(pAAV-CaMK        6768 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6768)
  AUTHORS   Institute for Basic Science
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     promoter        157..520
                     /label=CamKIIa promoter
                     /note="color: #993300; direction: RIGHT"
     primer_bind     546..608
                     /label=HA-CRY2(EA9,NheI)-F
                     /note="color: black; sequence: 
                     GACTGCTAGCGCCACCATGGGATACCCATACGACGTGCCTGACTACGCCCCACCGGTCA
                     TGAAGATGGAC; added: 2022-02-13"
     primer_bind     546..605
                     /label=FLAG-CRY2(EA9,NheI)-F
                     /note="color: black; sequence: 
                     GACTGCTAGCGCCACCATGGGAGACTACAAGGATGACGACGATAAGCCACCGGTCATGA
                     AGATG; added: 2022-02-13"
     CDS             join(558..563,564..587)
                     /codon_start=1
                     /label=FLAG tag
                     /note="This feature has 2 segments:
                        1: 558 .. 563 / #99ccff
                        2: 564 .. 587 / #99ccff"
                     /translation="MGDYKDDDDK"
     primer_bind     596..621
                     /label=CRY2(EA9,AgeI)-F
                     /note="color: black; sequence: 
                     GACTACCGGTCGCCACCATGAAGATGGACAAAAAGACCATCG; added: 
                     2021-08-12"
     CDS             597..2090
                     /codon_start=1
                     /label=CRY2(E281A)
                     /note="color: #33cccc"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGA
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     primer_bind     complement(1414..1445)
                     /label=Fragment 1.REV
                     /note="color: black; sequence: 
                     GCTTTCGGCGCCTTCGCTGTTTTTATCGCGAG"
     primer_bind     complement(1414..1445)
                     /label=STIM1(E281A)-R
                     /note="color: black; sequence: 
                     GCTTTCGGCGCCTTCGCTGTTTTTATCGCGAG"
     primer_bind     1431..1460
                     /label=Fragment 2.FOR
                     /note="color: black; sequence: 
                     GAAGGCGCCGAAAGCGCCGATCTGTTCCTG"
     primer_bind     1431..1460
                     /label=PHR(E281A)-F
                     /note="color: black; sequence: 
                     GAAGGCGCCGAAAGCGCCGATCTGTTCCTG"
     primer_bind     complement(1606..1624)
                     /label=HA-CRY2(EA9)-R
                     /note="color: black; sequence: GCATCCACGAGTGGGTACC; added: 
                     2022-02-13"
     primer_bind     complement(1672..1697)
                     /label=CRY2(internal)-R
                     /note="color: black; sequence: GACAGCAAATGAGGACACGATTACCC; 
                     added: 2022-02-21"
     CDS             2091..2117
                     /codon_start=1
                     /label=A9
                     /note="color: #ffffff"
                     /translation="ARDPPDLDN"
     CDS             2211..3554
                     /codon_start=1
                     /label=STIM1(238-685)
                     /note="color: #ff0000"
                     /translation="KEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHL
                     EKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSW
                     YAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSS
                     SLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPS
                     WMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTH
                     SDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPA
                     LAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRAL
                     QASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK"
     primer_bind     complement(2859..2879)
                     /label=STIM1(~460)-stop-BamHI-JGSR
                     /note="color: black; sequence: 
                     TCAGTTATCTAGATCCGGTGGATCCCTACCAGCTGGGGTCTATGTTGAG"
     primer_bind     3563..3603
                     /label=BamHI-XhoI-CW3SL-JGSF
                     /note="color: black; sequence: 
                     TGGATGGGCAGTTAGGGATCCctcgagataatcaacctctggattacaaaatttg"
     misc_feature    3578..4004
                     /label=W3SL
                     /note="Optimization of AAV expression cassettes to improve 
                     packaging capacity and transgene expression in neurons
                     WPRE3 + Late SV40
                     "
                     /note="color: #ffffff"
     primer_bind     complement(3631..3651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     primer_bind     complement(3993..4034)
                     /label=CW3SL-RsrII-JGSR
                     /note="color: black; sequence: 
                     tcctgcggccgctcggtccgtcctgcggccgctttaaaaaac"
     repeat_region   4031..4171
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4246..4701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4983..5087
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(5088..5156,5157..5948)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5088 .. 5156 / #ccffcc / signal sequence
                        2: 5157 .. 5948 / #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6119..6707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaactt gtggactaag tttgttcaca
      181 tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg
      241 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa
      301 agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc
      361 ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca
      421 cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag
      481 gagcacgggc aggcgagtgg cccctagttc tgggggcagc tctagagccg atccccgggt
      541 accgggctag cgccaccatg ggagactaca aggatgacga cgataagcca ccggtcatga
      601 agatggacaa aaagaccatc gtctggtttc ggagagattt gagaatagaa gataatcccg
      661 cgctcgccgc cgcggcccac gagggttccg tcttccccgt tttcatttgg tgtcctgaag
      721 aagaaggcca gttttatccc ggaagggcct ctaggtggtg gatgaagcaa agtctggccc
      781 atcttagcca gtcactgaaa gcactgggca gtgatcttac cctgatcaag acacacaata
      841 ccatctctgc catcctcgac tgcatcaggg tgaccggcgc aacgaaagtc gtgtttaacc
      901 acctgtacga tccagttagt ctggtgcgcg accacactgt gaaggagaag ctggtggaac
      961 gggggatcag tgtgcagagc tacaacgggg accttctgta cgagccatgg gagatctatt
     1021 gcgagaaagg gaaaccgttc acctccttca acagttactg gaagaaatgt ttggatatgt
     1081 caatagagtc cgttatgttg ccccctccct ggagactgat gccgattact gctgctgcag
     1141 aggccatctg ggcctgctcc atcgaggaac tcggtctgga aaatgaagca gaaaagccaa
     1201 gcaatgcact tctcactaga gcctggagcc ccggctggtc taatgccgac aagctgctta
     1261 acgagttcat cgaaaaacaa ctgattgact acgcgaagaa ctccaagaaa gtggtaggta
     1321 actcaactag cttgctctct ccatatctcc attttggcga gatttctgtc cgccatgtat
     1381 ttcagtgcgc tcggatgaaa cagattatct gggctcgcga taaaaacagc gaaggcgccg
     1441 aaagcgccga tctgttcctg cgagggatcg gacttcggga atactcccgg tatatatgtt
     1501 tcaactttcc attcacacac gagcagagtc tgttgtccca cctcaggttc ttcccctggg
     1561 acgccgatgt cgacaaattc aaggcatgga gacagggaag gacagggtac ccactcgtgg
     1621 atgctggcat gagagagctc tgggctacag gctggatgca caaccgcatc cgggtaatcg
     1681 tgtcctcatt tgctgtcaag tttctgctcc tgccttggaa atggggaatg aagtactttt
     1741 gggataccct tctcgacgcc gacttggagt gtgacattct gggatggcaa tatattagcg
     1801 ggtcaattcc tgacggccat gagttggaca ggttggacaa tccggccttg cagggagcta
     1861 agtatgatcc cgaaggagag tatattcgac agtggctccc cgagctggcc cgacttccta
     1921 cggagtggat tcaccatcct tgggacgcac cactgacagt gctcaaggca agcggggtgg
     1981 agctgggcac caattacgct aagcctatag ttgatataga tacagcacgc gagctgctgg
     2041 ctaaagcgat ctctcgcact cgggaggcgc agattatgat cggtgctgcc gccagggacc
     2101 cccccgacct cgacaacgaa ttcagtgggg gaggcggagg gggtggttca ggtggtggtg
     2161 gtggtggtgg ttcgggtggt ggcggagggg gaggctccgg actcagatct aaggagcaca
     2221 tgaagaagat gatgaaggac ttggaggggt tacaccgagc tgagcagagt ctgcatgacc
     2281 ttcaggaaag gctgcacaag gcccaggagg agcaccgcac agtggaggtg gagaaggtcc
     2341 atctggaaaa gaagctgcgc gatgagatca accttgctaa gcaggaagcc cagcggctga
     2401 aggagctgcg ggagggtact gagaatgagc ggagccgcca aaaatatgct gaggaggagt
     2461 tggagcaggt tcgggaggcc ttgaggaaag cagagaagga gctagaatct cacagctcat
     2521 ggtatgctcc agaggccctt cagaagtggc tgcagctgac acatgaggtg gaggtgcaat
     2581 attacaacat caagaagcaa aatgctgaga agcagctgct ggtggccaag gagggggctg
     2641 agaagataaa aaagaagaga aacacactct ttggcacctt ccacgtggcc cacagctctt
     2701 ccctggatga tgtagatcat aaaattctaa cagctaagca agcactgagc gaggtgacag
     2761 cagcattgcg ggagcgcctg caccgctggc aacagatcga gatcctctgt ggcttccaga
     2821 ttgtcaacaa ccctggcatc cactcactgg tggctgccct caacatagac cccagctgga
     2881 tgggcagtac acgccccaac cctgctcact tcatcatgac tgacgacgtg gatgacatgg
     2941 atgaggagat tgtgtctccc ttgtccatgc agtcccctag cctgcagagc agtgttcggc
     3001 agcgcctgac ggagccacag catggcctgg gatctcagag ggatttgacc cattccgatt
     3061 cggagtcctc cctccacatg agtgaccgcc agcgtgtggc ccccaaacct cctcagatga
     3121 gccgtgctgc agacgaggct ctcaatgcca tgacttccaa tggcagccac cggctgatcg
     3181 agggggtcca cccagggtct ctggtggaga aactgcctga cagccctgcc ctggccaaga
     3241 aggcattact ggcgctgaac catgggctgg acaaggccca cagcctgatg gagctgagcc
     3301 cctcagcccc acctggtggc tctccacatt tggattcttc ccgttctcac agccccagct
     3361 ccccagaccc agacacacca tctccagttg gggacagccg agccctgcaa gccagccgaa
     3421 acacacgcat tccccacctg gctggcaaga aggctgtggc tgaggaggat aatggctcta
     3481 ttggcgagga aacagactcc agcccaggcc ggaagaagtt tcctctcaaa atctttaaga
     3541 agcctcttaa gaagtaggaa ttgggatccc tcgagataat caacctctgg attacaaaat
     3601 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3661 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3721 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3781 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgtttattt gtgaaatttg
     3841 tgatgctatt gctttatttg taaccatcta gctttatttg tgaaatttgt gatgctattg
     3901 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3961 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcggccgc aggacggacc
     4021 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4081 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     4141 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     4201 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4261 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4321 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4381 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4441 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4501 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4561 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     4621 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4681 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4741 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4801 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4861 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4921 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4981 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5041 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5101 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5161 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5221 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5281 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5341 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5401 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5461 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5521 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5581 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5641 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5701 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5761 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5821 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5881 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5941 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6001 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6061 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6241 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6361 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6421 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6481 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6541 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6601 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6661 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6721 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
