LOCUS       sequence-198248-        5624 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV expression of ultrasensitive protein calcium sensor from 
            Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  AUTHORS   Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, 
            Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, 
            Kim DS
  TITLE     Ultrasensitive fluorescent proteins for imaging neuronal activity.
  JOURNAL   Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
  PUBMED    23868258
REFERENCE   2  (bases 1 to 5624)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2013-07-18- 18"; volume: "499"; issue: "7458"; 
            pages: "295-300"
COMMENT     Sequence Label: pAAV.Syn.GCaMP6m.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      102..111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(108..116,117..134,135..137,138..170,171..173,174..197,
                     198..224,225..281,282..287,288..557,558..575,576..1010,
                     1011..1016,1017..1460)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      204..213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     494..515
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(621..642)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(882..901)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1473..2061
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1526..1546)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1944..1955)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2064..2080)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2065..2081)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2110..2231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2121..2140)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2175..2194
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2393..2522
                     /label=AAV2 ITR
     rep_origin      complement(2759..3347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2839..2858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3518..4309,4310..4378))
                     /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     4141..4160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4379..4483)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4509..4964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4646..4667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4858..4877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5036..5165
                     /label=AAV2 ITR
     promoter        join(5247..5624,1..70)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
       61 gagagcgcag tcgaattcaa gctgctagca aggatccacc cgccaccatg ggttctcatc
      121 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
      181 acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt aagtggaata
      241 agacaggtca cgcagtcaga gctataggtc ggctgagctc actcgagaac gtctatatca
      301 aggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg
      361 acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
      421 tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa gaccccaacg
      481 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
      541 tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt
      601 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      661 gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      721 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      781 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      841 tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      901 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      961 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc
     1021 aactgactga agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg
     1081 gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc
     1141 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa
     1201 tcgacttccc tgagttcctg acaatgatgg caagaaaagg gagctacagg gacacggaag
     1261 aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag
     1321 cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg
     1381 aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac
     1441 aaatgatgac agcgaagtga aagcttatcg ataatcaacc tctggattac aaaatttgtg
     1501 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1561 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1621 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1681 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1741 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1801 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     1861 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     1921 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     1981 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2041 ccctttgggc cgcctccccg catcgatacc gtcgacctcg acccgggcgg ccgcttcgag
     2101 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2161 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2221 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     2281 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc
     2341 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     2401 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2461 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2521 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2581 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2641 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2701 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2761 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2821 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2881 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2941 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3001 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3061 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3121 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3181 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3241 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3301 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3361 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3421 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3481 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4381 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4441 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4501 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4561 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4621 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4681 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4741 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4801 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4861 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4921 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     4981 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg
     5041 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     5101 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     5161 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
     5221 gatctctgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
     5281 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
     5341 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
     5401 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
     5461 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
     5521 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
     5581 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcgg
//
