LOCUS       sequence-128106-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging vector that expresses ER-retained low-affinity 
            GCaMP3 variant under control of the SYN1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Henderson MJ, Baldwin HA, Werley CA, Boccardo S, Whitaker LR, Yan X,
            Holt GT, Schreiter ER, Looger LL, Cohen AE, Kim DS, Harvey BK
  TITLE     A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic 
            Reticulum Calcium Store.
  JOURNAL   PLoS One. 2015 Oct 9;10(10):e0139273. doi: 
            10.1371/journal.pone.0139273. eCollection 2015.
  PUBMED    26451944
REFERENCE   2  (bases 1 to 5993)
  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: "PLoS One.";
            date: "2015-10-9"; volume: "10(10):e0139273. doi"; pages: " 
            10.1371/journal.pone.0139273. eCollection 2015"
COMMENT     Sequence Label: pOTTC814 - pAAV-SYN1-CRTsigpep-GCaMP3
            (D324G+D...
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(714..722,723..740,741..743,744..776,777..779,780..803,
                     804..830,831..887,888..893,894..1163,1164..1181,1182..1616,
                     1617..1619,1620..1622,1623..2063)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKGGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDAGGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKGGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGGGQVNYEEFVQMMTAK"
     CDS             join(714..722,723..740,741..743,744..776,777..779,780..803,
                     804..830,831..887,888..893,894..1163,1164..1181,1182..1616,
                     1617..1619,1620..1622,1623..2063)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKGGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDAGGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKGGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGGGQVNYEEFVQMMTAK"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1100..1121
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1227..1248)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1488..1507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2120..2708
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2173..2193)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2591..2602)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2740..3216
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2939..2958)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3256..3396
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3256..3385
                     /label=AAV2 ITR
     rep_origin      3471..3926
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3558..3577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3768..3789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3943..3962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4062..4084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4122..4140)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4208..4312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4313..4381,4382..5173)
                     /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(4531..4550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5344..5932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5833..5852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat ccgctagcca
      661 ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccatgggtt
      721 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
      781 atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt
      841 ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc gagaacgtct
      901 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
      961 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     1021 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc
     1081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1141 tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg
     1201 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1261 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     1321 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1381 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1441 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact
     1501 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1561 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacacgc
     1621 gtgaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca
     1681 agggcgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc
     1741 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc ggcggtgacg
     1801 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa gacacagaca
     1861 gtgaagaaga aattagagaa gcgttccgtg tgtttgataa gggtggcaat ggctacatca
     1921 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     1981 ttgatgaaat gatcagggaa gcagacatcg atgggggtgg tcaggtaaac tacgaagagt
     2041 ttgtacaaat gatgacagcg aagacagcag aaaaagatga gttgtagggc gcgccgaatt
     2101 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2161 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2221 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2281 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2341 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2401 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2461 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2521 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     2581 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2641 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2701 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     2761 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     2821 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     2881 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     2941 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3001 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3061 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3121 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3181 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     3241 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3301 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3361 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3421 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3661 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3781 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3901 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3961 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4021 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4081 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4141 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4201 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4261 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4501 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4621 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4861 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4921 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5161 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
