LOCUS       sequence-406151-        7569 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Production of AAV particles encoding WHaloCaMP1a-EGFP from the 
            CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7569)
  AUTHORS   Farrants H, Shuai Y, Lemon WC, Hernandez CM, Yang S, Patel R, Qiao 
            G, Frei MS, Grimm JB, Hanson TL, Tomaska F, Turner GC, Stringer C, 
            Keller PJ, Beyene AG, Chen Y, Liang Y, Lavis LD, Schreiter ER
  TITLE     A modular chemigenetic calcium indicator enables in vivo functional 
            imaging with near-infrared light.
  JOURNAL   bioRxiv. 2023 Jul 19:2023.07.18.549527. doi: 
            10.1101/2023.07.18.549527. Preprint.
  PUBMED    37503182
REFERENCE   2  (bases 1 to 7569)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv. 
            2023 Jul 19:2023.07.18.549527. doi: 10.1101/2023.07.18.549527. 
            Preprint."
COMMENT     Sequence Label: pAAV-CaMKII-WHaloCaMP1a-EGFP
FEATURES             Location/Qualifiers
     source          1..7569
                     /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        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             2157..2597
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(2964..2966,2967..2969,2970..3680)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3696..4284
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3749..3769)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4167..4178)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4316..4792
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4515..4534)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4832..4972
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4832..4961
                     /label=AAV2 ITR
     rep_origin      5047..5502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5134..5153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5344..5365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5519..5538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5638..5660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5698..5716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5784..5888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5889..5957,5958..6749)
                     /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(6107..6126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6920..7508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7409..7428
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccgcc accatgctgc agaacgagct tgctcttaag ttggctggac
     1501 ttgatattaa caagactgga ggttctgcag aaatcggtac tggctttcca ttcgaccccc
     1561 attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca
     1621 cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc
     1681 cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg
     1741 acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg
     1801 aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt
     1861 tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc
     1921 gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc
     1981 gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagtggacgc
     2041 tgccgatggc cgtcgcgcgc gcccgtcgta aatggcagaa aacaggccat gcggttcgtg
     2101 ctatcggtcg tttgtcgagc ggaggctctg gcggttccgg cggttcaggt ggatcagatc
     2161 aattaacaga ggaacagatt gcggagttta aggaagcgtt ctctttattt gataaggatg
     2221 gcgacggtac aatcactact aaagaattgg gaacagtcat gcgctcattg gggcaaaatc
     2281 cgacagaggc tgaattgcag gacatgatta acgaggtaga cgccgatggg aacgggacta
     2341 tcgactttcc ggaatttctt actatgatgg cacgcaaaat gaaagatacc gattctgaag
     2401 aagagatccg tgaagccttc cgtgtttttg ataaagatgg gaacggctac atcagtgctg
     2461 ctgagttacg ccatgtgatg acaaatctgg gggaaaaact taccgacgaa gaagtagacg
     2521 aaatgattcg cgaggcggat attgacgggg atggacaagt aaactacgag gaatttgtgc
     2581 agatgatgac cgccaagtat ctgactgaag tcgagatgga ccattaccgc gagccgttcc
     2641 tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg
     2701 agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg
     2761 tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc
     2821 gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc
     2881 tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg
     2941 agatttccgg cggcggaagc ggcatggtct ctaaggggga ggaactcttt actggtgtgg
     3001 tgccaatact tgtagagctc gatggggacg ttaatggaca caaattttct gtctcaggtg
     3061 agggtgaggg agatgcaact tatggaaaac tgacattgaa attcatctgc acaacaggca
     3121 aattgcctgt cccctggcct accctggtaa ccaccctcac ttatggcgtt cagtgcttct
     3181 cccggtatcc tgatcacatg aaacagcatg atttcttcaa atcagcaatg cccgaaggtt
     3241 atgttcaaga gcggaccatc tttttcaagg acgatggaaa ctataaaacc agagctgagg
     3301 ttaagttcga gggagatact ttggtgaatc ggatcgaact gaaggggata gacttcaaag
     3361 aggatgggaa cattttgggg cataaactcg aatataacta caactcccac aacgtgtata
     3421 tcatggctga caaacagaag aacgggatca aagtaaattt taagatacga cacaatatag
     3481 aggacggtag tgtccaactt gctgatcact accagcagaa tacacccatc ggagacggac
     3541 ccgttctctt gcccgataat cactatctct ctacacaatc cgctctgtct aaagatccaa
     3601 acgaaaagcg ggaccacatg gtactgcttg agttcgttac agccgctggt atcaccttgg
     3661 gcatggatga gttgtataaa taaaagctta tcgataatca acctctggat tacaaaattt
     3721 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3781 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3841 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3901 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3961 agctcctttc cggaactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4021 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4081 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     4141 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4201 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4261 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     4321 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     4381 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     4441 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     4501 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     4561 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     4621 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     4681 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     4741 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     4801 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4861 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4921 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4981 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     5041 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5101 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     5161 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5221 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     5281 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5341 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     5401 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     5461 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5521 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5581 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5641 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5701 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5881 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6001 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6061 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6121 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6421 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7561 ctcacatgt
//
