LOCUS       sequence-310664-        7156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cell body-targeted GCaMP6f, under synapsin promoter: nullCoChR (the 
            optogenetic protein CoChR mutated to have 0 current), followed by 
            GCaMP6f and the KV2.1 motif (nullCoChR-GCaMP6f-Kv2.1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  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 7156)
  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: "Neuron. 
            2020 Jun 26. pii: S0896-6273(20)30398-6. doi: 
            10.1016/j.neuron.2020.05.029."
COMMENT     Sequence Label: pAAV-Syn-nullCoChR-GCaMP6f-Kv2.1
FEATURES             Location/Qualifiers
     source          1..7156
                     /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)"
     CDS             808..1671
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSANCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             join(1708..1716,1717..1734,1735..1737,1738..1770,1771..1773,
                     1774..1797,1798..1824,1825..1881,1882..1887,1888..2157,
                     2158..2175,2176..2610,2611..2616,2617..3057)
                     /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      1804..1813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2094..2115
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2221..2242)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2482..2501)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3289..3877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3342..3362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3760..3771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3897..4373
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4096..4115)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4419..4559
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4419..4548
                     /label=AAV2 ITR
     rep_origin      4634..5089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4721..4740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4931..4952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5106..5125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5225..5247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5285..5303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5371..5475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5476..5544,5545..6336)
                     /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(5694..5713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6507..7095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6996..7015
                     /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 ctggggaacg gcagcgccat tgtgcctatc
      841 gaccagtgct tttgcctggc ttggaccgac agcctgggaa gcgatacaga gcagctggtg
      901 gccaacatcc tccagtggtt cgccttcggc ttcagcatcc tgatcctgat gttctacgcc
      961 taccagactt ggagagccac ttgcggttgg gaggaggtct acgtctgttg cgtcgagctg
     1021 accaaggtca tcatcgagtt cttccacgag ttcgacgacc ccagcatgct gtacctggct
     1081 aacggacacc gagtccagtg gctgagatac gcagagtggc tgctgacttg tcccgtcatc
     1141 ctgatccacc tgagcaacct gaccggcctg aaggacgact acagcaagcg gaccatgagg
     1201 ctgctggtgt cagacgtggg aaccatcgtg tggggagcta caagcgccat gagcacaggc
     1261 tacgtcaagg tcatcttctt cgtgctgggt tgcatctacg gcgccaacac cttcttccac
     1321 gccgccaagg tgtatatcga gagctaccac gtggtgccaa agggcagacc tagaaccgtc
     1381 gtgcggatca tggcttggct gttcttcctg tcttggggca tgttccccgt gctgttcgtc
     1441 gtgggaccag aaggattcga cgccatcagc gtgtacggct ctaccattgg ccacaccatc
     1501 atcgacctca tgagcgctaa ttgttggggc ctgctgggac actatctgag agtgctgatc
     1561 caccagcaca tcatcatcta cggcgacatc cggaagaaga ccaagatcaa cgtggccggc
     1621 gaggagatgg aagtggagac catggtggac caggaggacg aggagacagt gggaggttca
     1681 ggtggaaccg gtggaagtgg aggtaccatg ggttctcatc atcatcatca tcatggtatg
     1741 gcttccatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
     1801 ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga
     1861 gctataggtc ggctgagttc actcgagaac gtctatatca aggccgacaa gcagaagaac
     1921 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1981 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2041 tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     2101 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     2161 ggaaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2221 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     2281 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2341 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2401 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     2461 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2521 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2581 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     2641 gcagaattta aagaggagtt ctccctattt gacaaggacg gggatgggac aataacaacc
     2701 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     2761 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     2821 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     2881 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     2941 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     3001 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagggt
     3061 gctagccaga gccagcctat cctgaacacc aaggagatgg cccctcagag caagcctcct
     3121 gaggagctgg agatgagcag catgcctagc cctgtggccc ctctgcctgc caggaccgag
     3181 ggcgtgatcg acatgaggag catgagcagc atcgacagct tcatcagctg cgccaccgac
     3241 ttccctgagg ccaccaggtt ctaagaattc cgtacaaagc ttatcgataa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccggct cgagacgggt
     3901 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3961 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     4021 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     4081 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     4141 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     4201 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     4261 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     4321 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctcgagc
     4381 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4441 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4501 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4561 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4621 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4681 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4741 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4801 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4861 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4921 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4981 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     5041 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5101 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5161 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5221 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5281 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5341 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5401 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5461 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5521 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5581 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5641 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5701 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5761 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5821 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5881 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5941 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6001 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6061 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6121 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6181 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6241 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6301 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6361 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6421 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6481 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6601 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6661 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6721 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6781 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6841 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6901 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6961 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7021 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7081 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7141 ccttttgctc acatgt
//
