LOCUS Exported 5657 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of jRCaMP1b, a red fluorescent calcium sensor protein, from the Synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV.Syn.NES-jRCaMP1b.WPRE.SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5657) 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 5657) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5657 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 341..358 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 362..394 /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 398..421 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" misc_feature 1673..2261 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1726..1746) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2144..2155) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2264..2280) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2265..2281) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2310..2431 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2321..2340) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2375..2394 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2593..2722 /label=AAV2 ITR primer_bind complement(2788..2805) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2959..3547) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3039..3058) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3718..4578) /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 4341..4360 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4579..4683) /gene="bla" /label=AmpR promoter rep_origin complement(4709..5164) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4846..4867) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5058..5077 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5236..5365 /label=AAV2 ITR promoter 5447..237 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact 61 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac 121 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca 181 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg 241 aattcaagct gctagcaagg atccacccgc caccatgctg cagaacgagc ttgctcttaa 301 gttggctgga cttgatatta acaagactgg aggaggttct catcatcatc atcatcatgg 361 tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa 421 ggatctcgca acaatggtcg actcatcgcg acgtaagtgg aataagtggg gtcacgcagt 481 cagagctata ggtcggctga gctcagcgaa caacaccgaa atgatgtacc cagcggatgg 541 tggtctgcgt ggttacactc acatggcgct gaaagttgat ggcggcggtc acctgtcctg 601 ttctttcgtg accacctacc gctccaaaaa gactgtcggc aacattaaga tgcctgccat 661 tcattacgtc agccaccgtc tggagcgcct ggaggagagc gataacgaaa tgtttgtcgt 721 acagcgtgaa cacgcagttg ccaagtttgt gggcctgggt ggtggcggcg gtaccggagg 781 gagcatgaac tccctgatca aggagaacat gcgtatgaaa gtggttctgg aaggctccgt 841 aaacggccac cagttcaaat gcactggtga aggcgaaggc aacccgtata tgggcaccca 901 gactatgcgt atcaaagtga tcgagggtgg tccgctgccg tttgcgttcg acatcctggc 961 gacgtccttt atgtatggct cccgtacctt catcaaatat ccgaaaggca tcccggattt 1021 ctttaagcag tccttcccgg aaggttttac ctgggaacgt gtgacccgtt acgaagacgg 1081 cggcgtaatt accgttatgc aagacacgtc tctggaggat ggctgcctgg tgtatcacgt 1141 gcaggttcgc ggtgtgaact tcccgagcaa tggtgctgta atgcaaaaga aaaccaaagg 1201 ttgggagcct acggactccc aactgactga agagcagatc gcagaattta aagaggcttt 1261 ctccctattt gacaaggacg gggatgggac aataacaacc aaggagatgg ggacggtgat 1321 gcggtctctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga 1381 tgccgacggt gacggcacaa tcgacttccc tgagttcctg attatgatgg caggcaaaat 1441 gaaatacaca gacagtgaag aagaaattag agaagcgttc ggcgtgtttg ataaggatgg 1501 caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt 1561 aacagatgaa gaggttgatg aaatgatcag ggaagcagac agcgatgggg atggtcaggt 1621 aaactacgaa gagtttgtac aaatgatgac agcgaagtga aagcttatcg ataatcaacc 1681 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 1741 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 1801 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 1861 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 1921 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 1981 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 2041 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 2101 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 2161 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 2221 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg 2281 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac 2341 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt 2401 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt 2461 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg 2521 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa 2581 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 2641 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 2701 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 2761 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 2821 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 2881 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 2941 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3001 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 3061 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 3121 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 3181 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3241 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3301 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 3361 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 3421 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 3481 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 3541 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 3601 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 3661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 3721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 3781 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 3841 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 3901 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 3961 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4021 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4081 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4141 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 4201 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 4261 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4321 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 4381 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 4441 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 4501 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 4561 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 4621 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 4681 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc 4741 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 4801 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4861 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 4921 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 4981 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 5041 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 5101 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 5161 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 5221 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 5281 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 5341 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct 5401 acgtagccat gctctaggaa gatctctgca gagggccctg cgtatgagtg caagtgggtt 5461 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg 5521 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa 5581 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc 5641 ccccgcctgg cggcgcg //