LOCUS Exported 6286 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cell body-targeted GCaMP6f, under synapsin 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 pAAV-Syn-SomaGCaMP6f2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6286) 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 6286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6286 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 58..187 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" polyA_signal 243..291 /note="synthetic polyadenylation signal" misc_feature 305..396 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 345..364 /label=RVprimer3 /note="pGL3 vector, forward primer" promoter 432..879 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 888..897 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 894..2243 /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 990..999 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 1280..1301 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1407..1428) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1668..1687) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 2622..3210 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2675..2695) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3093..3104) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3243..3260) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3249..3456 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3465..3594 /label=AAV2 ITR primer_bind complement(3613..3630) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3613..3629) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3622..3644) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3771..4226 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3858..3877) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4068..4089 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4293..4312) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4412..4434 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4472..4490) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4558..4662 /gene="bla" /label=AmpR promoter CDS 4663..5523 /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(4881..4900) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5694..6282 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6183..6202 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct 61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt 121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg 181 ggttcctgcg gccgcacgcg ttctagaaat attaaggtac gggaggtact tggagcggcc 241 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag 301 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg 361 tccccagtgc aagtgcaggt gccagaacat ttctctattc tagactcgag ctgcagaggg 421 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac 481 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat 541 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct 601 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc 661 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt 721 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg 781 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg 841 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cggatccgcc accatgggtt 901 ctcatcatca tcatcatcat ggtatggctt ccatgactgg tggacagcaa atgggtcggg 961 atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt 1021 ggaataagac aggtcacgca gtcagagcta taggtcggct gagttcactc gagaacgtct 1081 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca 1141 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg 1201 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc 1261 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 1321 tcggcatgga cgagctgtac aagggcggaa ccggagggag catggtgagc aagggcgagg 1381 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 1441 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt 1501 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 1561 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 1621 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact 1681 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 1741 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgc 1801 cggaccaact gactgaagag cagatcgcag aatttaaaga ggagttctcc ctatttgaca 1861 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc 1921 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg 1981 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca 2041 cggaagaaga aattagagaa gcgttcggtg tgtttgataa ggatggcaat ggctacatca 2101 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg 2161 ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt 2221 ttgtacaaat gatgacagcg aaggagctcg ggggcagcgg tgggtctgga gggactgggg 2281 gtagcggtgg tagcggtggg acagggggaa gcggcggtag cggtgggaca ctggaaatcg 2341 aggccgcttt cctggagcag gagaacaccg ccctggagac agaggtggct gagctggagc 2401 aggaggtgca gcggctggag aacatcgtgt ctcagtacga gaccagatac ggaccactgg 2461 gatccctgga gatccgggcc gcttttctga ggagacggaa caccgccctg aggacaaggg 2521 tggctgagct gaggcagaga gtgcagcggc tgcgcaacat cgtgagccag tacgagacac 2581 gctacggacc tctgtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca 2641 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 2701 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 2761 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 2821 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 2881 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 2941 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 3001 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga 3061 ttctgcgcgg gacgcccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 3121 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 3181 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgactagag ctcgctgatc 3241 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 3301 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 3361 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 3421 ggaggattgg gaagacaata gcaggcatgc tggggagaga tctgaggaac ccctagtgat 3481 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt 3541 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa 3601 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 3661 ccaacttaat cgccttgcag cacatccccc tttcgccagc ctggcgtaat agcgaagagg 3721 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg 3781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 3841 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 3901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 3961 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 4021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 4081 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 4141 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 4201 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc 4261 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 4321 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 4381 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 4441 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 4501 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 4561 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 4621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 4681 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 4741 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 4801 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 4861 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 4921 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 4981 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 5041 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 5101 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 5161 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 5221 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 5281 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 5341 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 5401 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 5461 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 5521 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 5581 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 5641 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 5701 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5761 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 5821 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 5881 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 5941 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 6001 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 6061 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 6121 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 6181 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 6241 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacg //