LOCUS       sequence-492108-        6416 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Bioluminescent reporter for calcium signaling in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6416)
  AUTHORS   Lambert GG, Crespo EL, Murphy J, Boassa D, Luong S, Hu J, Sprecher 
            B, Tree MO, Orcutt R, Heydari D, Bell AB, Torreblanca Zanca A, 
            Hakimi A, Lipscombe D, Moore CI, Hochgeschwender U, Shaner NC
  TITLE     CaBLAM! A high-contrast bioluminescent Ca (2+) indicator derived 
            from an engineered Oplophorus gracilirostris luciferase.
  JOURNAL   bioRxiv. 2023 Jun 26:2023.06.25.546478. doi: 
            10.1101/2023.06.25.546478. Preprint.
  PUBMED    37425712
REFERENCE   2  (bases 1 to 6416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv. 
            2023 Jun 26:2023.06.25.546478. doi: 10.1101/2023.06.25.546478. 
            Preprint."
COMMENT     Sequence Label: pAAV hSyn CaBLAM
FEATURES             Location/Qualifiers
     source          1..6416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   43..172
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        217..664
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      737..746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      998..1007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1907..1963
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             2537..2554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2585..3173
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2638..2658)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3056..3067)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3205..3681
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3404..3423)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3721..3861
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3721..3850
                     /label=AAV2 ITR
     rep_origin      3936..4391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4023..4042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4233..4254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4408..4427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4527..4549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4587..4605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4673..4777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4778..4846,4847..5638)
                     /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(4996..5015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5809..6397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6298..6317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
       61 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      121 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      181 acgcgtgtgt ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag
      241 gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc
      301 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
      361 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
      421 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
      481 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
      541 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
      601 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
      661 gcagtcgaga aggtaccgga tcctctagag tcgagctcac cggtggcgcg ccaatattat
      721 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gaggataaca tggcctctct
      781 cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt
      841 gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa
      901 gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca
      961 gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg
     1021 ataccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg
     1081 ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc
     1141 tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa
     1201 gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa
     1261 tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc
     1321 taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa
     1381 gaccgagctc aacttcaagg agtggcaaaa ggcctttacc ggatttgaag acttcgttgg
     1441 tgactgggaa cagaccgctg cttacaacct ggaccaggtt ctggaacagg gtggtgtttc
     1501 ttctgttctg cagaccctgg ctgtttctgt tacccctatc cagcgtatcg ttcgttctgg
     1561 tgaaaacggt ctgaagatcg acatccatgt tatcatccct tacgaaggtc tgtctgctga
     1621 ccagatggct catatcgaag aagttttcaa ggttgtttac cctgttgacg accatcattt
     1681 caaggttatc atggaatacg gtaccctggt tatcgacggt gttaccccta acatgctgaa
     1741 ctacttcggt cgtccttacg aaggtatcgc tgttttcgac ggtaagaaga tcaccgttac
     1801 cggtaccctg tggaacggta acaagatcat cgacgaacgt ctgatcaccc ctgagggttc
     1861 tatgctgttc cgtgttacca tcaactctgc ttcttctgac tcttctaggc gtaagtggaa
     1921 caagaccggt catgctgttc gtgctatcgg tcgtctgtct tctggtggat ccggtggtag
     1981 tggtggatcc ggaggttctg gtggatccaa cctgcctgac cagctgaccg aagaacagat
     2041 cgctgaattc aaggaagagt tctctctgtt cgataaagat ggagacggta ccatcaccac
     2101 caaggaactg ggtaccgtga tgcgttctct gggtcagaac cctaccgaag ctgaactgca
     2161 agacatgatc aacgaagttg acgctgacgg tgacggtacc atcgacttcc ctgaattcct
     2221 gaccatgatg gctcgtaaga tgaagtacag ggacaccgaa gaggaaatcc gtgaagcttt
     2281 cggcgttttc gacaaggacg gtatcggtta catctctgct gctgaactgc gtcatgttat
     2341 gaccaacctg ggtgaaaagc tgaccgacga agaagttgac gaaatgatcc gtgaagctga
     2401 catcgacggt gacggtcagg ttaactacga agaattcgtt cagatgatga ccgctaagtc
     2461 tcctgctgtt accggttacc gtctgctcga agaaatctct aacctcgagt aacatatgca
     2521 cgcgtcaatt gccggtcatc atcaccatca ccattgagtt taattcgata tcaagcttat
     2581 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2641 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2701 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2761 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2821 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2881 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2941 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3001 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3061 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3121 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3181 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     3241 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3301 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3361 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3421 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3481 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3541 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3601 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3661 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     3721 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3781 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3841 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3901 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4141 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4261 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4381 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4501 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4561 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4621 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggcct
//
