LOCUS Exported 5458 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV backbone, mEGFP-CREB donor for G-CREB with S133A mutation. ACCESSION . VERSION . KEYWORDS AAV-SYN_mEGFP-CREB (S133A) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5458) AUTHORS Laviv T, Scholl B, Parra-Bueno P, Foote B, Zhang C, Yan L, Hayano Y, Chu J, Yasuda R TITLE In Vivo Imaging of the Coupling between Neuronal and CREB Activity in the Mouse Brain. JOURNAL Neuron. 2019 Dec 17. pii: S0896-6273(19)31037-2. doi: 10.1016/j.neuron.2019.11.028. PUBMED 31883788 REFERENCE 2 (bases 1 to 5458) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5458 /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 210..657 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 682..691 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 688..1404 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(733..754) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(994..1013) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1341..1362 /label=EGFP-C /note="EGFP, forward primer" repeat_region 2721..2861 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2721..2850 /label=AAV2 ITR rep_origin 2936..3391 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3023..3042) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3233..3254 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3408..3427) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3527..3549 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3587..3605) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3673..3777 /gene="bla" /label=AmpR promoter CDS 3778..4638 /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(3996..4015) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4809..5397 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5298..5317 /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 gcgtattacc gccatgcatt agttattaat agtaatcaat 181 tacggggtct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg 241 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc 301 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc 361 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg 421 agccacagcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact 481 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac 541 gcgaggcgcg agataggggg gcgcggacgc gaccatctgc gctgcggcgc cggcgactca 601 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg 661 aattggctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg 721 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 781 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 841 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 901 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 961 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 1021 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 1081 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 1141 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 1201 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 1261 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac 1321 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 1381 ctcggcatgg acgagctgta caagcctacc cctaccccta ccatgaccat ggactctgga 1441 gcagacaacc agcagagtgg agatgctgct gtaacagaag ctgaaagtca acaaatgaca 1501 gttcaagccc agccacagat tgccacatta gcccaggtat ccatgccagc agctcatgcg 1561 acgtcatctg ctcccactgt aaccttagtg cagctgccca atgggcagac agtccaggtc 1621 catggggtca tccaggcggc ccagccatca gttattcagt ctccacaagt ccaaacagtt 1681 cagtcttcct gtaaggactt aaaaagactt ttctccggaa ctcagatttc aactattgca 1741 gaaagtgaag attcacagga gtctgtggat agtgtaactg attcccaaaa acgaagggaa 1801 atcctttcaa ggaggcctgc ctacaggaaa attttgaatg acttatcttc tgatgcacca 1861 ggggtgccaa ggattgaaga agaaaaatca gaagaagaga cttcagcccc tgccatcacc 1921 actgtaacag tgccaacccc gatttaccaa actagcagtg ggcagtatat tgccattacc 1981 cagggaggag caatacagct ggctaacaat ggtaccgatg gggtacaggg cctgcagaca 2041 ttaaccatga ccaatgcagc tgccactcag ccgggtacta ccattctaca atatgcacag 2101 accactgatg gacagcagat tctagtgccc agcaaccaag ttgttgttca agctgcctct 2161 ggtgatgtac aaacatacca gattcgcaca gcacccacta gcaccattgc ccctggagtt 2221 gttatggcgt cctccccagc acttcctaca cagcctgctg aagaagcagc acgaaagaga 2281 gaggttcgtc taatgaagaa cagggaagca gcaagagaat gtcgtagaaa gaagaaagaa 2341 tatgtgaaat gtttagagaa cagagtggca gtgcttgaaa accaaaacaa aacattgatt 2401 gaggagctaa aagcacttaa ggacctttac tgccacaagt cagattaaaa gcttgtggca 2461 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 2521 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa 2581 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga cccaaattgc 2641 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca 2701 cgtgtggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg 2761 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 2821 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt 2881 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 2941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3001 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3121 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 3181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3241 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg 3301 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3361 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 3421 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 3481 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 3541 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 3601 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 3661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3721 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 3781 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 3841 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 3901 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 3961 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 4021 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 4081 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 4141 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 4201 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 4261 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4321 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4381 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 4441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 4501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 4561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 4621 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 4681 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 4741 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 4801 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 4861 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 4921 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 4981 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5041 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5101 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5161 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 5221 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5281 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5341 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 5401 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt //