LOCUS Exported 8156 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Chemogenetics expression plasmid. ACCESSION . VERSION . KEYWORDS AAV CAMKII PSAM4 GlyR IRES EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8156) AUTHORS Magnus CJ, Lee PH, Bonaventura J, Zemla R, Gomez JL, Ramirez MH, Hu X, Galvan A, Basu J, Michaelides M, Sternson SM TITLE Ultrapotent chemogenetics for research and potential clinical applications. JOURNAL Science. 2019 Mar 14. pii: science.aav5282. doi: 10.1126/science.aav5282. PUBMED 30872534 REFERENCE 2 (bases 1 to 8156) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8156 /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 149..1437 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" intron 1460..1592 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 1575..1594 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 2944..3529 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3111..3128) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS 3542..4258 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3584..3605) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3845..3864) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4192..4213 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4283..4871 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4336..4356) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4754..4765) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4903..5379 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(5102..5121) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5419..5559 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5419..5548 /label=AAV2 ITR rep_origin 5634..6089 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5721..5740) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5931..5952 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6106..6125) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6225..6247 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6285..6303) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6371..6475 /gene="bla" /label=AmpR promoter CDS 6476..7336 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPKTLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6694..6713) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7507..8095 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7996..8015 /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg 181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc 241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa 301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag 361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca 421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt 481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc 541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca 601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt 661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg 721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag 781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt 841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca 901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg 961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt 1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg 1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag 1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa 1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa 1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc 1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt 1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct 1441 agagcggtac cggatccagg taagtatcaa ggttacaaga caggtttaag gagaccaata 1501 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct 1561 tactgacatc cactttgcct ttctctccac aggtgatgaa ttcgctagcg ccaccatgcg 1621 ctgttctcca ggcggcgtgt ggctcgccct ggctgcttcc cttctgcacg ttagcctgca 1681 gggtgagttc cagcgcaaac tgtataagga gcttgttaag aattataacc ccctggagcg 1741 gccggtcgca aatgattccc agccactgac agtgtacttc agcctctcct tgctgcagat 1801 catggacgtg gatgaaaaga accaggtgct gaccactaat atttggttgc agatgtcctg 1861 gaccgatcac tacttgcagt ggaatgtgag cgaataccca ggtgtaaaga ctgtaagatt 1921 ccctgacggc caaatctgga aaccagatat cctgctgtac aacagcgcag acgaaaggtt 1981 tgatgcaaca tttcacacca acgtgggagt caattcttca ggccactgcc tgtacctgcc 2041 ccctggaatc ttcaagtcct catgctatat cgacgtccgc tggtttccct tcgacgtcca 2101 gcactgcaaa ctcaaattcg ggagctggag ctacggcgga tggagcctgg atctgcaaat 2161 gcaggaggct gacatctctg gttacatccc gaatggggag tgggaccttg tgggaatccc 2221 cggtaaaaga agcgagcgat tttatgaatg ctgcaaagag cccttcccag atgtcacctt 2281 cacagtgacc atgcggagac gcatgggtta ttatctgatc caaatgtata tcccaagctt 2341 gcttatagtg attttgtcat ggatctcctt ctggattaat atggacgccg ctccagctag 2401 ggtcggactg ggcatcacca cagtgctgac aatgactact cagagctcag gcagccgagc 2461 cagcttgccc aaggtttctt acgtgaaggc catcgatatc tggatggctg tctgccttct 2521 gtttgtcttc agcgcactgc tggaatacgc cgctgtcaat tttgtgtctc gacagcataa 2581 agagctgttg cggttcagaa gaaaacgacg ccaccacaaa gaggatgagg caggagaagg 2641 acgcttcaac tttagcgcct atggtatggg acctgcttgc ctccaggcta aagacggaat 2701 ttccgtgaag ggagccaaca atagcaacac aaccaaccca ccccctgctc catctaagag 2761 cccggaggaa atgcgcaaac tctttattca gagagcgaaa aagatcgaca aaatctcccg 2821 gatcggattc cccatggctt tcctgatttt caacatgttt tattggatca tctacaagat 2881 tgtgcgaagg gaggacgtac acaaccagta agcggccgca attccccccs ccvccccccc 2941 ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg 3001 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc 3061 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa 3121 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag 3181 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg 3241 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg 3301 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctaag cgtattcaac 3361 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg 3421 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac 3481 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tgggagatcc 3541 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 3601 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 3661 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 3721 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 3781 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 3841 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 3901 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 3961 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg 4021 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga 4081 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 4141 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 4201 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaac 4261 cggtgtcgac aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac 4321 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 4381 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 4441 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 4501 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 4561 aactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 4621 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 4681 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt 4741 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 4801 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 4861 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac 4921 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc 4981 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg 5041 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg 5101 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc 5161 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc 5221 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc 5281 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg 5341 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg 5401 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg 5461 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg 5521 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc 5581 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc 5641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 5701 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 5761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 5821 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc 5881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 5941 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat 6001 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 6061 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga 6121 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 6181 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 6241 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct 6301 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg 6361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 6421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 6481 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 6541 tgctcaccca aaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 6601 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 6661 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 6721 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 6781 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 6841 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 6901 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 6961 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 7021 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 7081 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 7141 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 7201 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 7261 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 7321 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 7381 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 7441 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 7501 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 7561 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 7621 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 7681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 7741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 7801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 7861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 7921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 7981 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 8041 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 8101 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt //