LOCUS Exported 5490 bp ds-DNA circular SYN 27-NOV-2019 DEFINITION AAV backbone, mEGFP-CREB donor for G-CREB. ACCESSION . VERSION . KEYWORDS AAV-SYN_mEGFP-CREB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5490) TITLE In vivo imaging of the coupling between neuronal and CREB activity in the mouse brain JOURNAL Unpublished REFERENCE 2 (bases 1 to 5490) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 27, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5490 /dnas_title="addgene-plasmid-137006 UPDATE" /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 /dnas_title="enhanced GFP" /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH MVLLEFVTAAGITLGMDELYK" 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 2753..2893 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2753..2882 /label=AAV2 ITR rep_origin 2968..3423 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3055..3074) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3265..3286 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3440..3459) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3559..3581 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3619..3637) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3705..3809 /dnas_title="bla" /gene="bla" /label=AmpR promoter CDS 3810..4670 /dnas_title="bla" /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" primer_bind complement(4028..4047) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4841..5429 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5330..5349 /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 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact 481 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac 541 gcgaggcgcg agataggggg gcacgggcgc 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 ggaggccttc 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 caacctgtag 2641 ggcctgcggg gtctattggg aacccaaatt gctgggatta caggcgtgaa ccactgctcc 2701 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc 2761 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga 2821 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 2881 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 2941 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg 3001 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3061 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3121 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3181 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3241 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 3301 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 3361 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa 3421 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 3481 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 3541 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 3601 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 3661 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 3721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 4021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 4081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 4141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 4201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 4261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 4321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 4381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 4441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 4501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 4561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 4621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 4681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 4741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 4801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 4861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 4921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 4981 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 5041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 5101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 5161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 5221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 5281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 5341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 5401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 5461 tggccttttg ctggcctttt gctcacatgt //