LOCUS       sequence-306415-        6436 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cell body-targeted GCaMP6f, under CAG promoter: GCaMP6f followed by 
            a linker, and the EE-RR coiled coil motif (GCaMP6f-27-EE-RR). As 
            bright as conventional GCaMP6f..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  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 6436)
  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-CAG-SomaGCaMP6f2
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     promoter        487..764
                     /label=chicken beta-actin promoter
     primer_bind     1044..1066
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1128..1147
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1186..1194,1195..1212,1213..1215,1216..1248,1249..1251,
                     1252..1275,1276..1302,1303..1359,1360..1365,1366..1635,
                     1636..1653,1654..2088,2089..2094,2095..2535)
                     /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      1282..1291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1572..1593
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1699..1720)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1960..1979)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2914..3502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2967..2987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3385..3396)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3545..3666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3556..3575)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3610..3629
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3828..3957
                     /label=AAV2 ITR
     rep_origin      complement(4194..4782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4274..4293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4953..5744,5745..5813))
                     /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     5576..5595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5814..5918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5944..6399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6081..6102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6293..6312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      181 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaggtt aattaaccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      541 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      601 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      661 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      721 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
      781 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      841 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      901 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      961 cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg acggctgcct
     1021 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1081 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1141 tattgtgctg tctcatcatt ttggcaaaga attggatccg ccaccatggg ttctcatcat
     1201 catcatcatc atggtatggc ttccatgact ggtggacagc aaatgggtcg ggatctgtac
     1261 gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa gtggaataag
     1321 acaggtcacg cagtcagagc tataggtcgg ctgagttcac tcgagaacgt ctatatcaag
     1381 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
     1441 ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1501 ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag
     1561 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1621 gacgagctgt acaagggcgg aaccggaggg agcatggtga gcaagggcga ggagctgttc
     1681 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1741 gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1801 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1861 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1921 cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1981 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2041 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct gccggaccaa
     2101 ctgactgaag agcagatcgc agaatttaaa gaggagttct ccctatttga caaggacggg
     2161 gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc
     2221 acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc
     2281 gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga cacggaagaa
     2341 gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat cagtgcagca
     2401 gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa
     2461 atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa
     2521 atgatgacag cgaaggagct cgggggcagc ggtgggtctg gagggactgg gggtagcggt
     2581 ggtagcggtg ggacaggggg aagcggcggt agcggtggga cactggaaat cgaggccgct
     2641 ttcctggagc aggagaacac cgccctggag acagaggtgg ctgagctgga gcaggaggtg
     2701 cagcggctgg agaacatcgt gtctcagtac gagaccagat acggaccact gggatccctg
     2761 gagatccggg ccgcttttct gaggagacgg aacaccgccc tgaggacaag ggtggctgag
     2821 ctgaggcaga gagtgcagcg gctgcgcaac atcgtgagcc agtacgagac acgctacgga
     2881 cctctgtaag aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2941 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3001 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3061 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3121 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3181 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3241 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3301 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3361 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3421 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3481 tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac
     3541 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3601 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3661 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3721 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga
     3781 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     3841 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3901 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3961 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4021 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4081 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4141 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4201 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4261 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4321 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4381 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4441 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4501 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4561 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4621 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4681 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4741 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4801 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4861 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4921 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4981 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5041 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgcgaccca
     5101 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5161 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5221 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5281 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5341 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5401 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5461 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5521 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5581 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5641 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5701 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5761 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5821 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5881 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5941 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6001 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6061 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6121 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6181 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6241 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6301 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6361 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     6421 gcaactgttg ggaagg
//
