LOCUS Exported 6462 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For AAV based expression of nuclear jRGECO1a in neurons in Cre-negative neurons with FLEx switch. ACCESSION . VERSION . KEYWORDS SLF155: AAV-syn-DO-H2B-jRGECO1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6462) AUTHORS Farhi SL, Parot VJ, Grama A, Yamagata M, Abdelfattah AS, Adam Y, Lou S, Jun Kim J, Campbell RE, Cox DD, Cohen AE TITLE Wide-area all-optical neurophysiology in acute brain slices. JOURNAL J Neurosci. 2019 Apr 5. pii: JNEUROSCI.0168-19.2019. doi: 10.1523/JNEUROSCI.0168-19.2019. PUBMED 30952812 REFERENCE 2 (bases 1 to 6462) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6462 /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 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 660..693 /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 744..777 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 791..1168 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" regulatory 1181..1190 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1202..1258 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKAGHAVRAIGRLSS" primer_bind 1954..1973 /label=mCherry-F /note="mCherry, forward primer" protein_bind complement(2447..2480) /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(2531..2564) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2589..3177 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2642..2662) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3060..3071) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3209..3685 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3408..3427) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3725..3865 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3725..3854 /label=AAV2 ITR rep_origin 3940..4395 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4027..4046) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4237..4258 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4412..4431) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4531..4553 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4591..4609) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4677..4781 /gene="bla" /label=AmpR promoter CDS 4782..5642 /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(5000..5019) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5813..6401 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6302..6321 /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa 661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc 721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc 781 gcgccgcacc atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa 841 ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag 901 ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc 961 caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga 1021 ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac 1081 ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa 1141 ggccatcacc aagtacacca gcgctaagga tccaccggtc gccaccatgg tcgactcatc 1201 acgtcgtaag tggaataagg caggtcacgc agtcagagct ataggtcggc tgagctcacc 1261 cgtggtttcc gagcggatgt accccgagga cggcgccctg aagagcgaga tcaagaaggg 1321 gctgaggctg aaggacggcg gccactacgc cgccgaggtc aagaccacct acaaggccaa 1381 gaagcccgtg cagctgcccg gcgcctacat cgtggacatc aagttggaca tcgtgtccca 1441 caacgaggac tacaccatcg tggaacagtg cgaacgcgcc gagggccgcc actccaccgg 1501 cggcatggac gagctgtaca agggaggtac aggcgggagt ctggtgagca agggcgagga 1561 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt 1621 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca 1681 gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc 1741 ccctcagttc atgtacggct ccaaggccta cattaagcac ccagccgaca tccccgacta 1801 cttcaagctg tccttccccg agggcttcag gtgggagcgc gtgatgaact tcgaggacgg 1861 cggcattatt cacgtgaacc aggactcctc cctgcaggac ggcgtattca tctacaaggt 1921 gaagctgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg 1981 ctgggaggct acgcgtgacg acctgactga agagcagatc gcagaattta aagaggcttt 2041 ctccctattt gacaaggacg gggatgggac gataacaacc aaggagcttg ggacggtgtt 2101 ccgatcgctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga 2161 tgccgacggt gacggcacat tcgacttccc tgagttcctg acgatgatgg caagaaaaat 2221 gaatgacaca gacagtgaag aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg 2281 caatggctac atcggcgcag cagagcttcg ccacgtgatg acagaccttg gagagaagtt 2341 aacagatgag gaggttgatg aaatgatcag ggtagcagac atcgatgggg atggtcaggt 2401 aaactacgaa gagtttgtac aaatgatgac agcgaagtag gctagcataa cttcgtataa 2461 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat 2521 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc 2581 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 2641 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 2701 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 2761 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 2821 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 2881 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 2941 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 3001 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 3061 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 3121 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc 3181 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc 3241 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag 3301 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt 3361 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc 3421 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc 3481 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag 3541 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa 3601 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg 3661 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg 3721 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact 3781 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc 3841 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg 3901 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat 3961 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag 4021 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 4081 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 4141 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt 4201 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 4261 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg 4321 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 4381 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 4441 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 4501 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 4561 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta 4621 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg 4681 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 4741 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 4801 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 4861 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 4921 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 4981 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 5041 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 5101 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 5161 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 5221 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 5281 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 5341 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 5401 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 5461 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 5521 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 5581 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 5641 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 5701 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 5761 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 5821 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 5881 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 5941 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 6001 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 6061 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 6121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 6181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 6241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 6301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 6361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 6421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt //