LOCUS       sequence-404981-        6487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of AAV particles encoding WHaloCaMP1a-EGFP from the 
            synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  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 6487)
  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: "bioRxiv. 
            2023 Jul 19:2023.07.18.549527. doi: 10.1101/2023.07.18.549527. 
            Preprint."
COMMENT     Sequence Label: pAAV-synapsin-WHaloCaMP1a-EGFP
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             863..1303
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1670..1672,1673..1675,1676..2386)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2402..2990
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2455..2475)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2873..2884)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2993..3009)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2994..3010)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3039..3160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3050..3069)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3104..3123
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3322..3451
                     /label=AAV2 ITR
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4447..5238,5239..5307))
                     /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     5070..5089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5308..5412)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5438..5893)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5575..5596)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5787..5806
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5965..6094
                     /label=AAV2 ITR
     promoter        join(6176..6487,1..136)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
       61 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      121 gtgcctgaga gcgcagtcga attcaagctg ctagcaagga tccgccacca tgctgcagaa
      181 cgagcttgct cttaagttgg ctggacttga tattaacaag actggaggtt ctgcagaaat
      241 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
      301 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
      361 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
      421 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
      481 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
      541 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
      601 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
      661 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
      721 tcagaacgtt tttatcgagt ggacgctgcc gatggccgtc gcgcgcgccc gtcgtaaatg
      781 gcagaaaaca ggccatgcgg ttcgtgctat cggtcgtttg tcgagcggag gctctggcgg
      841 ttccggcggt tcaggtggat cagatcaatt aacagaggaa cagattgcgg agtttaagga
      901 agcgttctct ttatttgata aggatggcga cggtacaatc actactaaag aattgggaac
      961 agtcatgcgc tcattggggc aaaatccgac agaggctgaa ttgcaggaca tgattaacga
     1021 ggtagacgcc gatgggaacg ggactatcga ctttccggaa tttcttacta tgatggcacg
     1081 caaaatgaaa gataccgatt ctgaagaaga gatccgtgaa gccttccgtg tttttgataa
     1141 agatgggaac ggctacatca gtgctgctga gttacgccat gtgatgacaa atctggggga
     1201 aaaacttacc gacgaagaag tagacgaaat gattcgcgag gcggatattg acggggatgg
     1261 acaagtaaac tacgaggaat ttgtgcagat gatgaccgcc aagtatctga ctgaagtcga
     1321 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1381 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1441 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1501 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1561 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1621 gatcgcgcgc tggctgtcga cgctcgagat ttccggcggc ggaagcggca tggtctctaa
     1681 gggggaggaa ctctttactg gtgtggtgcc aatacttgta gagctcgatg gggacgttaa
     1741 tggacacaaa ttttctgtct caggtgaggg tgagggagat gcaacttatg gaaaactgac
     1801 attgaaattc atctgcacaa caggcaaatt gcctgtcccc tggcctaccc tggtaaccac
     1861 cctcacttat ggcgttcagt gcttctcccg gtatcctgat cacatgaaac agcatgattt
     1921 cttcaaatca gcaatgcccg aaggttatgt tcaagagcgg accatctttt tcaaggacga
     1981 tggaaactat aaaaccagag ctgaggttaa gttcgaggga gatactttgg tgaatcggat
     2041 cgaactgaag gggatagact tcaaagagga tgggaacatt ttggggcata aactcgaata
     2101 taactacaac tcccacaacg tgtatatcat ggctgacaaa cagaagaacg ggatcaaagt
     2161 aaattttaag atacgacaca atatagagga cggtagtgtc caacttgctg atcactacca
     2221 gcagaataca cccatcggag acggacccgt tctcttgccc gataatcact atctctctac
     2281 acaatccgct ctgtctaaag atccaaacga aaagcgggac cacatggtac tgcttgagtt
     2341 cgttacagcc gctggtatca ccttgggcat ggatgagttg tataaataaa agcttatcga
     2401 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2461 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2521 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2581 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2641 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2701 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2761 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2821 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2881 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2941 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3001 tcgacctcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3061 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3181 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     3241 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     3301 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3361 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3421 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     3481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5401 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5461 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5521 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5581 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5641 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5701 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5761 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5821 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5881 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5941 cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     6001 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     6061 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
     6121 tacttatcta cgtagccatg ctctaggaag atctctgcag agggccctgc gtatgagtgc
     6181 aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg
     6241 acccactgga caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg
     6301 ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag
     6361 tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga
     6421 cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg
     6481 ccgccgg
//
