LOCUS Exported 5684 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of jRGECO1a, a red fluorescent calcium sensor protein, from the Synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV.Syn.NES-jRGECO1a.WPRE.SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5684) AUTHORS Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G, Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS TITLE Sensitive red protein calcium indicators for imaging neural activity. JOURNAL Elife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727. PUBMED 27011354 REFERENCE 2 (bases 1 to 5684) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5684 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 354..371 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 375..407 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 411..434 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 462..518 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKAGHAVRAIGRLSS" primer_bind 1214..1233 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1713..2301 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1766..1786) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2184..2195) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2304..2320) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2305..2321) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2350..2471 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2361..2380) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2415..2434 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2633..2762 /label=AAV2 ITR primer_bind complement(2828..2845) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2999..3587) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3079..3098) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3758..4618) /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 4381..4400 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4619..4723) /gene="bla" /label=AmpR promoter rep_origin complement(4749..5204) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4886..4907) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5098..5117 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5276..5405 /label=AAV2 ITR promoter 5487..250 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg 61 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc 121 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg 181 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct 241 gagagcgcag tcgaattcaa gctgctagca aggatccacc cgccaccatg ctgcagaacg 301 agcttgctct taagttggct ggacttgata ttaacaagac tggaggaggt tctcatcatc 361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg 421 acgatgacga taaggatctc gcaacaatgg tcgactcatc acgtcgtaag tggaataagg 481 caggtcacgc agtcagagct ataggtcggc tgagctcacc cgtggtttcc gagcggatgt 541 accccgagga cggcgccctg aagagcgaga tcaagaaggg gctgaggctg aaggacggcg 601 gccactacgc cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 661 gcgcctacat cgtggacatc aagttggaca tcgtgtccca caacgaggac tacaccatcg 721 tggaacagtg cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 781 agggaggtac aggcgggagt ctggtgagca agggcgagga ggataacatg gccatcatca 841 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga 901 tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag ctgaaggtga 961 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct 1021 ccaaggccta cattaagcac ccagccgaca tccccgacta cttcaagctg tccttccccg 1081 agggcttcag gtgggagcgc gtgatgaact tcgaggacgg cggcattatt cacgtgaacc 1141 aggactcctc cctgcaggac ggcgtattca tctacaaggt gaagctgcgc ggcaccaact 1201 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggct acgcgtgacg 1261 acctgactga agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg 1321 gggatgggac gataacaacc aaggagcttg ggacggtgtt ccgatcgctg gggcagaacc 1381 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacat 1441 tcgacttccc tgagttcctg acgatgatgg caagaaaaat gaatgacaca gacagtgaag 1501 aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg caatggctac atcggcgcag 1561 cagagcttcg ccacgtgatg acagaccttg gagagaagtt aacagatgag gaggttgatg 1621 aaatgatcag ggtagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac 1681 aaatgatgac agcgaagtga aagcttatcg ataatcaacc tctggattac aaaatttgtg 1741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 1801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 1861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 1921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 1981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 2041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 2101 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 2161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 2221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 2281 ccctttgggc cgcctccccg catcgatacc gtcgacctcg acccgggcgg ccgcttcgag 2341 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa 2401 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca 2461 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt 2521 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc 2581 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc 2641 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga 2701 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 2761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 3421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 4741 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 4801 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 4861 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 4921 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 4981 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 5041 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 5101 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 5161 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag 5221 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg 5281 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg 5341 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg 5401 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa 5461 gatctctgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg 5521 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt 5581 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg 5641 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc cccc //