LOCUS Exported 5849 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of ultrafast protein calcium sensor under the Syn promoter, Cre-dependent expression. ACCESSION . VERSION . KEYWORDS pGP-AAV-syn-FLEX-jGCaMP8f-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5849) AUTHORS Zhang , Yan; Rozsa, Marton; Bushey, Daniel; Zheng, Jihong; Reep, Daniel; Broussard, Gerard Joey; Tsang, Arthur; Tsegaye, Getahun; Patel, Ronak; Narayan, Sujatha; Li, Jing Xuan; Zhang, Rongwei; Ahrens, Misha B.; Turner, Glenn C.; Wang, Samuel S.-H.; Svoboda, Karel; Korff, Wyatt; Schreiter, Eric R.; Hasseman, Jeremy P.; Kolb, Ilya; Looger, Loren L TITLE jGCaMP8 Fast Genetically Encoded Calcium Indicators JOURNAL Janelia Research Campus. Online resource. 2020 REFERENCE 2 (bases 1 to 5849) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5849 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 34..481 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind complement(511..544) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(607..640) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 1210..1229 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1469..1490 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1596..1617) /label=EGFP-C /note="EGFP, forward primer" CDS complement(1899..1916) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" protein_bind 1936..1969 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 2040..2073 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2115..2703 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2168..2188) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2586..2597) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2746..2867 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2757..2776) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2811..2830 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3029..3158 /label=AAV2 ITR primer_bind complement(3224..3241) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3395..3983) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3475..3494) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4154..5014) /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 4777..4796 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5015..5119) /gene="bla" /label=AmpR promoter rep_origin complement(5145..5600) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5282..5303) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5494..5513 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 5672..5801 /label=AAV2 ITR ORIGIN 1 ctaggaagat ctctgcagag ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat 61 gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac 121 ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg 181 aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg 241 cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca 301 aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca 361 ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga 421 ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca 481 gtcgaattca agctgctagc aaggatcccc ataacttcgt ataaagtatc ctatacgaag 541 ttatatcaaa ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat 601 cgacccataa cttcgtataa tgtatgctat acgaagttat actagagctt tcacttcgct 661 gtcatcattt gtacaaactc ttcgtagttt acctgaccat ccccatcgat gtctgcttcc 721 ctgatcattt catcaacctc ttcatctgtt aacttctctc caaggtttgt catcacgtgg 781 cgaagctctg ctgcactgat gtagccattg ccatccttat caaacacacc gaacgcttct 841 ctaatttctt cttccgtgtc cctgtatttc atttttcttg ccatcattgt caggaactca 901 gggaagtcga ttgtgccgtc accgtcggca tctacttcat tgatcatgtc ctgcagctct 961 gcttctgtgg ggttcagtcc gagagaccgc atcaccgtcc ccagctcctt ggttgttatt 1021 gtcccatccc cgtccttgtc aaatagggag aaagcctctt taaattctgc gatctgctct 1081 tcagtcagtt ggtccggcag gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc 1141 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc 1201 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggatgtag 1261 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg 1321 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg 1381 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcaccctcg 1441 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc 1501 accccggtga acagctcctc gcccttgctc accatgctcc ctccggtacc gcccttgtac 1561 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga 1621 tcgcgcttct cgttggggtc tttcgaaagt ttggactcca cgctcaggta gtggttgtcg 1681 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggta ggcgagctgc 1741 acgccgccgt cctcgatgtt gtggcggatg tggaagttcg ccttgatgcc gttcttctgc 1801 ttgtcggcct tgatatagac gttgatcttc aggcccatca ggcgggcgat gatcttcacg 1861 gcggtggcca cctccttgaa ggtcttcttg cgacgcgtgt gatgatggtg atgatgcatg 1921 gtggcggatc ctagtataac ttcgtatagg atactttata cgaagttatc attgggattc 1981 ttcctatttt gatccaagca tcaccatcga ccctctagtc cagatctcac catcgaccca 2041 taacttcgta tagcatacat tatacgaagt tatgtccctc gaagaggttc gaattcgata 2101 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 2161 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 2221 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 2281 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 2341 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 2401 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 2461 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 2521 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 2581 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 2641 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 2701 cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg 2761 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 2821 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 2881 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa 2941 acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag 3001 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc 3061 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc 3121 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac 3181 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 3241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 3301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 4441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 4741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 4861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 4921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 4981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 5041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 5101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa 5161 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc 5221 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt 5281 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa 5341 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg 5401 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg 5461 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc 5521 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa 5581 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg 5641 atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca ctgaggccgc 5701 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc 5761 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc 5821 gccatgctac ttatctacgt agccatgct //