LOCUS       sequence-310657-        7360 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cell body-targeted GCaMP6f under synapsin promoter: GCaMP6f followed
            by a linker, the tail region of AnkyrinG, and an ER export motif 
            (GCaMP6f-27-AnkTail-motif-ER2). Not as bright as SomaGCaMP6f2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7360)
  AUTHORS   Shemesh OA, Linghu C, Piatkevich KD, Goodwin D, Celiker OT, Gritton 
            HJ, Romano MF, Gao R, Yu CJ, Tseng HA, Bensussen S, Narayan S, Yang 
            CT, Freifeld L, Siciliano CA, Gupta I, Wang J, Pak N, Yoon YG, 
            Ullmann JFP, Guner-Ataman B, Noamany H, Sheinkopf ZR, Park WM, Asano
            S, Keating AE, Trimmer JS, Reimer J, Tolias AS, Bear MF, Tye KM, Han
            X, Ahrens MB, Boyden ES
  TITLE     Precision Calcium Imaging of Dense Neural Populations via a 
            Cell-Body-Targeted Calcium Indicator.
  JOURNAL   Neuron. 2020 Jun 26. pii: S0896-6273(20)30398-6. doi: 
            10.1016/j.neuron.2020.05.029.
  PUBMED    32592656
REFERENCE   2  (bases 1 to 7360)
  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"; journalName: "Neuron. 
            2020 Jun 26. pii: S0896-6273(20)30398-6. doi: 
            10.1016/j.neuron.2020.05.029."
COMMENT     Sequence Label: pAAV-Syn-SomaGCaMP6f1
FEATURES             Location/Qualifiers
     source          1..7360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        341..788
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      802..811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(808..816,817..834,835..837,838..870,871..873,874..897,
                     898..924,925..981,982..987,988..1257,1258..1275,1276..1710,
                     1711..1716,1717..2157)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      904..913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1194..1215
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1321..1342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1582..1601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3493..4081
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3546..3566)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3964..3975)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4101..4577
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4300..4319)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4623..4763
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4623..4752
                     /label=AAV2 ITR
     rep_origin      4838..5293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4925..4944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5135..5156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5310..5329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5429..5451
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5489..5507)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5575..5679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5680..5748,5749..6540)
                     /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(5898..5917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6711..7299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7200..7219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aatggttaat taatctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
      361 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
      421 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      481 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      541 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      601 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      661 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      721 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      781 gagcgcagtc gagtaggatc cgccaccatg ggttctcatc atcatcatca tcatggtatg
      841 gcttccatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
      901 ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
      961 gctataggtc ggctgagttc actcgagaac gtctatatca aggccgacaa gcagaagaac
     1021 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1081 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1141 tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     1201 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     1261 ggaaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1321 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     1381 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1441 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1501 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     1561 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1621 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1681 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     1741 gcagaattta aagaggagtt ctccctattt gacaaggacg gggatgggac aataacaacc
     1801 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     1861 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     1921 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     1981 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     2041 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     2101 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaaggag
     2161 ctcggcggat ctggcggtag cggagggaca ggtggctctg gagggagcgg aggcactggc
     2221 ggatctggtg ggagcggagg taccagagaa gggagaatag acgatgaaga acctttcaaa
     2281 atcgttgaga aagtgaagga agacttagtc aaagtcagtg agatactcaa aaaggacgtg
     2341 tgtgtcgaga gcaaagggcc acccaagtcc ccgaagagtg acaaaggaca ctctccagag
     2401 gatgactgga cagaatttag ttctgaggaa atacgagaag ccaggcaggc tgcggccagt
     2461 cacgccccgt ccctgcctga gagagtacac gggaaggcca acctcactag agtcatagac
     2521 tacctgacca atgacatcgg gagcagctca ctgaccaact taaagtacaa gtttgaagag
     2581 gctaagaagg atggagagga gagacagaag aggattttaa agccagccat ggccttgcag
     2641 gagcataaac tcaaaatgcc gccggcctcc atgaggcctt ccacctctga gaaggaactg
     2701 tgcaagatgg ctgactcttt ttttggagca gatgccatcc tagagtctcc cgatgacttt
     2761 tcccagcatg accaagataa aagtcccttg tctgacagtg gctttgaaac ccgcagtgag
     2821 aaaacacctt cggccccgca aagtgccgag agcacaggcc ctaagccgtt atttcacgaa
     2881 gtccccatcc cccctgtcat cacagagaca agaactgaag tggtccatgt gatcaggagc
     2941 tatgagccct cttctggaga gatcccccag tcccagccag aggaccccgt gtctcccaag
     3001 ccttccccta cttttatgga gttggaacca aaacccacca cctctagcat caaggaaaaa
     3061 gtaaaagcat ttcagatgaa agccagtagt gaggaagagg accacagtcg ggttctaagc
     3121 aaaggcatgc gtgtcaaaga agagactcac atcaccacga ccaccaggat ggtgtatcac
     3181 tcccctcctg gtggcgagtg cgcctccgaa aggattgagg agaccatgtc agtccatgac
     3241 atcatgaagg cctttcagtc cgggcgagac ccttccaaag agctggcagg gctgtttgaa
     3301 cacaagtcgg ccatgtctcc agatgttgcc aagtctgctg ctgagacctc agcccagcac
     3361 gcagagaagg acagccaaat gaaacccaaa ctggagcgca ttatagaagt gcacatcgaa
     3421 aaaggtccgc agagtccttg tgagttctgc tacgagaatg aagtgtaaga attccgtaca
     3481 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3541 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3601 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3661 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3721 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3781 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3841 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3901 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3961 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4021 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4081 catcgatacc ggctcgagac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     4141 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     4201 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     4261 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     4321 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     4381 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     4441 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     4501 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     4561 ctcccttccc tgtccttctc gagctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     4621 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4681 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4741 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4801 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5041 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5161 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5281 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5341 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5401 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5461 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5521 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5581 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5641 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5701 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5761 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5821 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5881 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5941 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6001 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6061 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6121 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6181 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6241 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6301 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6361 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6421 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6481 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6541 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6601 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7321 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
