LOCUS       sequence-267844-        6096 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, slow variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  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 6096)
  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-jYCaMP1s
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   52..181
                     /label=AAV2 ITR
     intron          774..906
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     889..908
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    919..952
                     /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    1015..1048
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1625..1644
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1884..1905
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2011..2032)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2245..2301)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRVIGRLSS"
     regulatory      2313..2322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2347..2380)
                     /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(2451..2484)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2556..3144
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2609..2629)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3027..3038)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3287..3306
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3505..3634
                     /label=AAV2 ITR
     rep_origin      3706..4161
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3793..3812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4003..4024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4187..4291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4292..4360,4361..5152)
                     /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(4510..4529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5323..5911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5812..5831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgcgcgct
       61 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
      121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      181 ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc
      241 cgagctccag tgtgctggaa ttcgcccttt tcagtattta aattgacgac cgaccccgac
      301 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      361 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      421 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      481 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      541 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      601 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      661 gtgtcgtgcc tgagagcgca gctgtgctcc tgggcaccgc gcagtccgcc cccgcggctc
      721 ctggccagac cacccctagg accccctgcc ccaagtcgca gccgttggat caggtaagta
      781 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa
      841 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
      901 ccacaggtga tgatccccat aacttcgtat aaagtatcct atacgaagtt atatcaaaat
      961 aggaagacca atgcttcacc atcgacccga attgccaagc atcaccatcg acacataact
     1021 tcgtataatg tatgctatac gaagttatac tagtgctagc ggatcctact tcgctgtcat
     1081 catttgtaca aactcttcgt agtttacctg accatcccca tcgatgtctg cttccctgat
     1141 catttcatca acctcttcat ctgttaactt ctctccaagg tttgtcatca cgtggcgaag
     1201 ctctgctgca ctgatgtagc cattgccatc cttatcaaac acaccgaacg cttctctaat
     1261 ttcttcttcc gtgtccctgt atttcatttt tcttgccatc attgtcagga actcagggaa
     1321 gtcgattgtg ccgtcaccgt cggcatctac ttcattgatc atgtcctgca gctctgcttc
     1381 tgtggggttc tgccccagag accgcatcac cgtccccagc tccttggttg ttattgtccc
     1441 atccccgtcc ttgtcaaata gggagaaaag ctctttaaat tctgcgatct gctcttcagt
     1501 cagttggtca ggaaggttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1561 gtcgatgccc ttaagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1621 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgca cgtagccttc
     1681 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     1741 ctgcaggccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1801 ggtgcaaatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcac cctcgccgga
     1861 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1921 ggtgaacagc tcctcgccct tgctcaccat gctccctccg gtaccgccct tgtacagctc
     1981 atccatgccc agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     2041 cttctcgttg gggtctttgc taagcacggg ctggtagctc aggtagtggt tgtcgggcag
     2101 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tgataggcga gctgcacgcc
     2161 gccgtcctcg atgttgtggc ggatgtggaa gttcgccttg atgccgttct tctgcttgtc
     2221 ggcggtgata tagacgttct cgagtgagct cagccgacct atcactctga ctgcgtgacc
     2281 tgtcttattc cacttacgac gtgatgagtc gaccatggtg gcgatatcac cggtggtacc
     2341 actagtataa cttcgtatag gatactttat acgaagttat cattgggatt cttcctattt
     2401 tgatccaagc atcaccatcg accctctagt ccaaatctca ccatcgaccc ataacttcgt
     2461 atagcataca ttatacgaag ttatgtccct cgaagaggtt cgaaacactg aaaagggcca
     2521 attctgcaaa tgatccactc gagaagctta tcgataatca acctctggat tacaaaattt
     2581 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2641 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2701 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2761 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2821 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2881 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2941 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3001 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3061 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3121 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg ctcgagcatg
     3181 catctagagt cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3241 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3301 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3361 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3421 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3481 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3541 tgcgcgctcg ctcgctcact gaggacgggc gaccaaaggt cgcccgacgc ccgggctttg
     3601 cccgggcggc ctcagtgagc gagcgagcgc gcagctggcg taatagcgaa gaggcccgca
     3661 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     3721 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3781 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3841 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3901 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3961 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4021 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4081 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4141 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4201 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4261 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4321 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4381 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4441 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4501 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4561 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4621 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4681 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4741 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4801 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4861 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4921 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4981 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5041 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5101 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5161 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5221 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5461 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5581 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5641 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5701 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5761 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5821 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5881 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5941 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6001 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6061 gcgcagcgag tcagtgagcg aggaagcgga agagcg
//
