LOCUS Exported 6575 bp ds-DNA circular SYN 21-SEP-2021 DEFINITION AAV transfer plasmid for neuronal expression of soma-targeted (RL-10, ribosomal tag) jGCaMP8s, with a flexible GS-linker sequence attaching the ribosomal tag to the jGCaMP8s protein.. ACCESSION . VERSION . KEYWORDS AAV-hSyn-RiboL1-jGCaMP8s SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6575) AUTHORS Grødem S, Nymoen I, Vatne GH, Bjornsdottir V, Lensjø KK, Fyhn M TITLE An updated suite of viral vectors for in vivo calcium imaging using local and retro-orbital injections JOURNAL bioRxiv REFERENCE 2 (bases 1 to 6575) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6575 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 183..630 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 667..684 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind 966..987 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1093..1114) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1354..1373) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 2702..3290 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2755..2775) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3173..3184) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3322..3798 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3521..3540) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3838..3978 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3838..3967 /label=AAV2 ITR rep_origin 4053..4508 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4140..4159) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4350..4371 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4525..4544) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4644..4666 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4704..4722) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4790..4894 /gene="bla" /label=AmpR promoter CDS 4895..5755 /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(5113..5132) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5926..6514 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6415..6434 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtttaatt aagtgtctag actgcagagg gccctgcgta 181 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc 241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca 301 gagaggggga ggggaaacag gatgcggcga ggctcgtgcg cactgccagc ttcagcaccg 361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc 421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc 481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg 541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc 601 ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga tccacccgcc 661 accatgcatc atcaccatca tcacacgcgt cgcaagaaga ccttcaagga ggtggccacc 721 gccgtgaaga tcatcgccat gctgatgggc ctgaagatca acgtctatat caaggccgac 781 aagcagaaga acggcatcaa ggcgaacttc cacatccgcc acaacatcga ggacggcggc 841 gtgcagctcg cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 901 cccgacaacc actacctgag cgtggagtcc aaactttcga aagaccccaa cgagaagcgc 961 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1021 ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg 1081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 1141 ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 1201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 1261 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 1321 ggctacatcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 1381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 1441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acctgccgga ccaactgact 1501 gaagagcaga tcgcagaata caaagaggct ttctccctat ttgacaagga cggggatggg 1561 acaataacaa ccaaggagct ggggacggtg atgcggtctc tcggacacaa ccccacagaa 1621 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc 1681 cctgagttcc tgacaatgat ggcaagaaaa atgaaataca gggacacgga agaagaaatt 1741 agagaagcgt tcggtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt 1801 cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc 1861 agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg 1921 acagcgaagg ggggcagcgg tgggtctgga gggactgggg gtagcggtgg tagcggtggg 1981 acagggggaa gcggcggtag tggtggaaca agcagcaaag tctcacgcga caccctgtac 2041 gaggcggtgc gggaagtcct gcacgggaac cagcgcaagc gccgcaagtt tctggagacg 2101 gtggagctgc agatcagcct gaagaactac gaccctcaga aggacaaacg tttctcgggc 2161 accgtcaggc tcaagtccac cccacgcccc aagttctcgg tgtgcgttct gggggaccag 2221 cagcactgtg atgaagccaa ggccgtggat atcccccaca tggacatcga ggcgctcaag 2281 aaacttaaca aaaacaagaa gttggtcaag aagctggcta agaagtacga tgcctttttg 2341 gcctctgagt ctctgattaa gcagatccca cgtatcctgg gcccaggcct aaacaaggct 2401 ggcaagttcc cctccctgct gacacacaat gaaaacatgg tggccaaagt ggatgaggtg 2461 aaatcgacaa tcaagttcca gatgaagaag gtgctgtgtt tggccgtcgc tgttggccac 2521 gtgaagatga ccgatgatga gctagtctac aacattcatc tggctgtcaa tttcttggtg 2581 tccttgctta agaaaaactg gcaaaacgtg cgggctctgt acatcaagag caccatgggc 2641 aagccccagc gtctgtatta gggcgcgccc ctgcagggaa ttcgatatca agcttatcga 2701 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 3001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 3061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 3121 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 3301 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg 3361 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc 3421 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc 3481 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg 3541 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc 3601 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt 3661 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa 3721 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact 3781 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg 3841 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 3901 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 3961 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 4021 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc 4081 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc 4141 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 4201 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 4261 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 4321 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 4381 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg 4441 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 4501 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc 4561 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 4621 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 4681 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 4741 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 4801 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 4861 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 4921 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 4981 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 5041 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 5101 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 5161 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 5221 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 5281 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 5341 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 5401 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 5461 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 5521 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 5581 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 5641 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 5701 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 5761 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 5821 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 5881 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 5941 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 6001 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 6061 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 6121 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 6181 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 6241 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 6301 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 6361 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 6421 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 6481 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 6541 gttcctggcc ttttgctggc cttttgctca catgt //