LOCUS Exported 5510 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses EGFP and bReaChES under a minimal CamKII Promoter. ACCESSION . VERSION . KEYWORDS pAAV-CamKII-GFP-P2A-bReaches-minWPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5510) AUTHORS Bernal Sierra YA, Rost BR, Pofahl M, Fernandes AM, Kopton RA, Moser S, Holtkamp D, Masala N, Beed P, Tukker JJ, Oldani S, Bonigk W, Kohl P, Baier H, Schneider-Warme F, Hegemann P, Beck H, Seifert R, Schmitz D TITLE Potassium channel-based optogenetic silencing. JOURNAL Nat Commun. 2018 Nov 5;9(1):4611. doi: 10.1038/s41467-018-07038-8. PUBMED 30397200 REFERENCE 2 (bases 1 to 5510) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5510 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 550..559 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 556..1272 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(601..622) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(862..881) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1209..1230 /label=EGFP-C /note="EGFP, forward primer" CDS 1288..1344 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" primer_bind complement(2393..2413) /label=WPRE-R /note="WPRE, reverse primer" repeat_region 2773..2913 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2773..2902 /label=AAV2 ITR rep_origin 2988..3443 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3075..3094) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3285..3306 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3460..3479) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3579..3601 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3639..3657) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3725..3829 /gene="bla" /label=AmpR promoter CDS 3830..4690 /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(4048..4067) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4861..5449 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5350..5369 /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 gcggccgcag atgctgcaca cttgtggact aagtttgttc gcatcccctt 181 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca 241 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa 301 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg 361 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc 421 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg 481 ggcaggcgag tggcccctag ttctgggggc agcgtgaaaa gacggtatcg ataagcttga 541 ttcgagctag ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc 1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1261 gagctgtaca aaccgcgggg atccggagct actaacttca gcctgctgaa gcaagcagga 1321 gatgttgaag aaaaccccgg gcctatggac tatggcggcg ctttgtctgc cgtcggactg 1381 ttccagacca gctacaccct ggagaacaac ggcagcgtga tctgcatccc caacaacggc 1441 cagtgcttct gcctggcctg gctgaagagt aacggcacca acgccgagaa gctggccgcc 1501 aacatcctgc agtgggtggt gtttgcgctg agcgtggcgt gcctgggctg gtatgcgtat 1561 caggcgtggc gcgcgacctg cggctgggaa gaagtgtatg tggcgctgat tgaaatgatg 1621 aaaagcatta ttgaagcgtt tcatgaattt gatagcccgg cgaccctgtg gctgagcagc 1681 ggcaacggcg tggtgtggat gcgctatggc tcatggctgc tgacctgccc cgtgattctg 1741 attcatctga gcaacctgac cggcctgaaa gatgattata gcaaacgcac catgggcctg 1801 ctggtgagcg acgtgggctg cattgtgtgg ggcgcgacca gcgcgatgtg caccggctgg 1861 accaaaattc tgttttttct gattagcctg agctatggca tgtataccta ttttcatgcg 1921 gccaaagtgt atattgaagc gtttcatacc gtgccgaaag gcctgtgcag acagctggtg 1981 agagccatgg cctggctgtt cttcgtgagc tggggcatgt tccccgtgct gttcctgctg 2041 ggccccgagg gcttcggcca tattagcccg tatggcagcg cgattggcca tagcattctg 2101 gatctgattg cgaagaacat gtggggcgtg ctgggcaact atctgcgcgt gaaaattcat 2161 gaacatattc tgctgtatgg cgatattcgc aaaaaacaga aaattaccat tgcgggccag 2221 gaaatggaag tggaaaccct ggtggcggaa gaagaagatg ggtcgaccaa gagcaggatc 2281 accagcgagg gcgagtacat ccccctggac cagatcgaca tcaacgtgta actcgagata 2341 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 2401 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 2461 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca 2521 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 2581 tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt 2641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt 2701 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt 2761 taaagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 2821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 2881 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta 2941 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag 3001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 3061 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 3121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 3181 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata 3241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 3301 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc 3361 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 3421 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc 3481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 3721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 3781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 3841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 3901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 3961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 4021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 4081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 4141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 4201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 4261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 4321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 4381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 4441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 4501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 4561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 4621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 4681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt //