LOCUS       sequence-267930-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yellow protein calcium sensor expressed under human Synapsin1 
            promoter, Cre mediated flip-excision switch, for AAV production or 
            direct transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  AUTHORS   Mohr MA, Bushey D, Aggarwal A, Marvin JS, Kim JJ, Marquez EJ, Liang 
            Y, Patel R, Macklin JJ, Lee CY, Tsang A, Tsegaye G, Ahrens AM, Chen 
            JL, Kim DS, Wong AM, Looger LL, Schreiter ER, Podgorski K
  TITLE     jYCaMP: an optimized calcium indicator for two-photon imaging at 
            fiber laser wavelengths.
  JOURNAL   Nat Methods. 2020 May 25. pii: 10.1038/s41592-020-0835-7. doi: 
            10.1038/s41592-020-0835-7.
  PUBMED    32451475
REFERENCE   2  (bases 1 to 6074)
  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: "Nat 
            Methods. 2020 May 25. pii: 10.1038/s41592-020-0835-7. doi: 
            10.1038/s41592-020-0835-7."
COMMENT     Sequence Label: pAAV-syn-FLEX-jYCaMP1
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          723..855
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     838..857
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    868..901
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    964..997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1574..1593
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1833..1854
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1960..1981)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2194..2250)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      2262..2271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2296..2329)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2400..2433)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2505..3093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2558..2578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2976..2987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3171..3292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3182..3201)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3236..3255
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3798..4386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3878..3897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4557..5348,5349..5417))
                     /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     5180..5199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5418..5522)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5548..6003)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5685..5706)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5897..5916
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcc gagctccagt gtgctggaat tcgccctttt cagtatttaa attgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag ctgtgctcct gggcaccgcg cagtccgccc
      661 ccgcggctcc tggccagacc acccctagga ccccctgccc caagtcgcag ccgttggatc
      721 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
      781 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
      841 cctttctctc cacaggtgat gatccccata acttcgtata aagtatccta tacgaagtta
      901 tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca tcaccatcga
      961 cacataactt cgtataatgt atgctatacg aagttatact agtgctagcg gatcctactt
     1021 cgctgtcatc atttgtacaa actcttcgta gtttacctga ccatccccat cgatgtctgc
     1081 ttccctgatc atttcatcaa cctcttcatc tgttaacttc tctccaaggt ttgtcatcac
     1141 gtggcgaagc tctgctgcac tgatgtagcc attgccatcc ttatcaaaca caccgaacgc
     1201 ttctctaatt tcttcttccg tgtccctgta tttcattttt cttgcgtaca ttgtcaggaa
     1261 ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact tcattgatca tgtcctgcag
     1321 ctctgcttct gtggggttct gccccagaga ccgcatcacc gtccccagct ccttggttgt
     1381 tattgtccca tccccgtcct tgtcaaatag ggagaaaagc tctttaaatt ctgcgatctg
     1441 ctcttcagtc agttggtcag gtgggttgta ctccagcttg tgccccagga tgttgccgtc
     1501 ctccttgaag tcgatgccct taagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1561 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctgcac
     1621 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1681 ggcgaagcac tgcaggccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag
     1741 cttgccggtg gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcacc
     1801 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1861 caccaccccg gtgaacagct cctcgccctt gctcaccatg ctccctccgg taccgccctt
     1921 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     1981 gtgatcgcgc ttctcgttgg ggtctttcga aagcacgggc tggtagctca ggtagtggtt
     2041 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag
     2101 ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga tgccgttctt
     2161 ctgcttgtcg gcggtgatat agacgttctc gagtgagctc agccgaccta tagctctgac
     2221 tgcgtgacct gtcttattcc acttacgacg tgatgagtcg accatggtgg cgatatcacc
     2281 ggtggtacca ctagtataac ttcgtatagg atactttata cgaagttatc attgggattc
     2341 ttcctatttt gatccaagca tcaccatcga ccctctagtc caaatctcac catcgaccca
     2401 taacttcgta tagcatacat tatacgaagt tatgtccctc gaagaggttc gaaacactga
     2461 aaagggccaa ttctgcaaat gatccactcg agaagcttat cgataatcaa cctctggatt
     2521 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2581 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2641 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2701 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2761 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2821 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2881 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     2941 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3001 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3061 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc
     3121 tcgagcatgc atctagagtc gacccgggcg gccgcttcga gcagacatga taagatacat
     3181 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3241 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3301 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     3361 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     3421 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3481 ctccctctct gcgcgctcgc tcgctcactg aggacgcccg ggctttgccc gggcggcctc
     3541 agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3601 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3661 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3841 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3901 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3961 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4021 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4141 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4201 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4261 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4321 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4381 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4501 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5401 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5521 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5581 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5641 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5701 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5761 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5821 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5881 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5941 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6001 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6061 cgctattacg ccag
//
