LOCUS Exported 6081 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION EGFP under the control of CaMKIIa promoter. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6081) AUTHORS Bryan Roth TITLE Roth lab DREADDs JOURNAL Unpublished REFERENCE 2 (bases 1 to 6081) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6081 /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" regulatory 1458..1467 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1464..2180 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1509..1530) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1770..1789) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2117..2138 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2208..2796 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2261..2281) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2679..2690) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2828..3304 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3027..3046) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3344..3484 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3344..3473 /label=AAV2 ITR rep_origin 3559..4014 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3646..3665) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3856..3877 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4031..4050) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4150..4172 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4210..4228) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4296..4400 /gene="bla" /label=AmpR promoter CDS 4401..5261 /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(4619..4638) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5432..6020 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5921..5940 /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 cggatccgcc accatggtga gcaagggcga ggagctgttc accggggtgg 1501 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1561 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 1621 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 1681 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 1741 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 1801 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1861 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1921 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1981 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 2041 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 2101 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 2161 gcatggacga gctgtacaag taggaattcg atatcaagct tatcgataat caacctctgg 2221 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 2281 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 2341 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 2401 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 2461 ccaccacctg tcagctcctt tccggaactt tcgctttccc cctccctatt gccacggcgg 2521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 2581 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca 2641 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 2701 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 2761 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga 2821 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc 2881 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta 2941 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg 3001 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca 3061 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc 3121 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta 3181 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta 3241 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt 3301 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg 3361 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc 3421 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg 3481 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 3541 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 3601 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct 3661 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 3721 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg 3781 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 3841 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg 3901 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc 3961 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt 4021 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 4081 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 4141 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 4201 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 4261 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 4321 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 4381 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 4441 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 4501 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 4561 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 4621 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 4681 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 4741 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 4801 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 4861 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 4921 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 4981 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 5041 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 5101 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 5161 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 5221 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 5281 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 5341 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 5401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 5461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 5521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 5581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 5641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 5701 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 5761 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 5821 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 5881 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 5941 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 6001 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 6061 gctggccttt tgctcacatg t //