LOCUS       sequence-268929-        6255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Axon-targeted 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 6255)
  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 6255)
  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-axon-jYCaMP1s
FEATURES             Location/Qualifiers
     source          1..6255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   39..168
                     /label=AAV2 ITR
     intron          761..893
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     876..895
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    906..939
                     /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    1002..1035
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1053..1496,1497..1502,1503..1937,1938..1955,
                     1956..2225,2226..2231,2232..2288,2289..2315,2316..2339,
                     2340..2342,2343..2375,2376..2378,2379..2396,2397..2402))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="GSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNKT
                     GHAVRVIGRLSSLENVYITADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSYQPVLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGY
                     GLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRSL
                     GQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGNG
                     YISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1612..1631
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1871..1892
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1998..2019)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2300..2309
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2493..2526)
                     /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(2597..2630)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2702..3290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2755..2775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3173..3184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3379..3398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3433..3452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4017..4605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4097..4116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4776..5567,5568..5636))
                     /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     5399..5418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5637..5741)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5767..6222)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5904..5925)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6116..6135
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
       61 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      121 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
      181 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatccga gctccagtgt
      241 gctggaattc gcccttttca gtatttaaat tgacgaccga ccccgaccca ctggacaagc
      301 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      361 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      421 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      481 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      541 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      601 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      661 gagcgcagct gtgctcctgg gcaccgcgca gtccgccccc gcggctcctg gccagaccac
      721 ccctaggacc ccctgcccca agtcgcagcc gttggatcag gtaagtatca aggttacaag
      781 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
      841 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgatga
      901 tccccataac ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg
      961 cttcaccatc gacccgaatt gccaagcatc accatcgaca cataacttcg tataatgtat
     1021 gctatacgaa gttatactag tgctagcgga tcctacttcg ctgtcatcat ttgtacaaac
     1081 tcttcgtagt ttacctgacc atccccatcg atgtctgctt ccctgatcat ttcatcaacc
     1141 tcttcatctg ttaacttctc tccaaggttt gtcatcacgt ggcgaagctc tgctgcactg
     1201 atgtagccat tgccatcctt atcaaacaca ccgaacgctt ctctaatttc ttcttccgtg
     1261 tccctgtatt tcatttttct tgccatcatt gtcaggaact cagggaagtc gattgtgccg
     1321 tcaccgtcgg catctacttc attgatcatg tcctgcagct ctgcttctgt ggggttctgc
     1381 cccagagacc gcatcaccgt ccccagctcc ttggttgtta ttgtcccatc cccgtccttg
     1441 tcaaataggg agaaaagctc tttaaattct gcgatctgct cttcagtcag ttggtcagga
     1501 aggttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgccctta
     1561 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     1621 ttgccgtcgt ccttgaagaa gatggtgcgc tcctgcacgt agccttcggg catggcggac
     1681 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg caggccgtag
     1741 cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcaaatcagc
     1801 ttcagggtca gcttgccgta ggtggcatcg ccctcaccct cgccggacac gctgaacttg
     1861 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     1921 tcgcccttgc tcaccatgct ccctccggta ccgcccttgt acagctcatc catgcccaga
     1981 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     2041 tctttgctaa gcacgggctg gtagctcagg tagtggttgt cgggcagcag cacggggccg
     2101 tcgccgatgg gggtgttctg ctggtagtga taggcgagct gcacgccgcc gtcctcgatg
     2161 ttgtggcgga tgtggaagtt cgccttgatg ccgttcttct gcttgtcggc ggtgatatag
     2221 acgttctcga gtgagctcag ccgacctatc actctgactg cgtgacctgt cttattccac
     2281 ttacgacgtg atgagtcgac catggtggcg agatccttat cgtcatcgtc gtacagatcc
     2341 cgacccattt gctgtccacc agtcatgcta gccataccgt gatgatgatg gtgatgagaa
     2401 ccaatctttt ggtcctcatc attcttttca acctgtttgg ttcttctcat acagcacagc
     2461 atggtggcga tatcaccggt ggtaccacta gtataacttc gtataggata ctttatacga
     2521 agttatcatt gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccaa
     2581 atctcaccat cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa
     2641 gaggttcgaa acactgaaaa gggccaattc tgcaaatgat ccactcgaga agcttatcga
     2701 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3121 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3301 tcgacccggg cggccgctcg agcatgcatc tagagtcgac ccgggcggcc gcttcgagca
     3361 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3421 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3481 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3541 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     3601 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     3661 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg acgcccgggc
     3721 tttgcccggg cggcgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4441 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5761 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5821 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5881 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5941 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     6001 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     6061 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     6121 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     6181 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     6241 tgcgcaactg ttggg
//
