LOCUS Exported 5446 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV plasmid with SYN1 promoter driving expression of EmGFP. Contains WPRE.. ACCESSION . VERSION . KEYWORDS pEMS2155 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5446) AUTHORS Galvan A, Petkau TL, Hill AM, Korecki AJ, Lu G, Choi D, Rahman K, Simpson E, Leavitt BR, Smith Y TITLE Intracerebroventricular Administration of AAV9-PHP.B SYN1-EmGFP Induces Widespread Transgene Expression in the Mouse and Monkey CNS. JOURNAL Hum Gene Ther. 2021 Apr 16. doi: 10.1089/hum.2020.301. PUBMED 33860682 REFERENCE 2 (bases 1 to 5446) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5446 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..448 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" intron 483..615 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 598..617 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 636..645 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 642..1361 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(687..708) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(948..967) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1295..1316 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1427..1447) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1845..1856) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1977..2098 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1988..2007) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2042..2061 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2267..2396 /label=AAV2 ITR primer_bind complement(2415..2432) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2415..2431) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2424..2446) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2573..3028 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2660..2679) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2870..2891 /label=F1ori-F /note="F1 origin, forward primer" promoter 3054..3158 /gene="bla" /label=AmpR promoter CDS 3159..4019 /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(3377..3396) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4190..4778 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4679..4698 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4932..4949 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5066..5087 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5102..5132 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5140..5156 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5145..5167 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5164..5180 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5164..5180 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 5218..5347 /label=AAV2 ITR ORIGIN 1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 421 aggagtcgtg tcgtgcctga gagcgcaggg cgcgccgaag ttggtcgtga ggcactgggc 481 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga 541 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg 601 cctttctctc cacaggtgtc cactcccagg cggccgccac catggtgagc aagggcgagg 661 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 721 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 781 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcacct 841 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 901 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 961 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 1021 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 1081 acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca 1141 agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 1201 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg 1261 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 1321 ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgcg atcgctcaac 1381 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 1441 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 1501 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 1561 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 1621 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 1681 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 1741 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg 1801 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 1861 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 1921 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgatcgcag acatgataag 1981 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg 2041 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 2101 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta 2161 aagcaagtaa aacctctaca aatgtggtag tcgaaattcc cgataaggat cttcctagag 2221 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg 2281 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag 2341 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt 2401 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt 2461 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag 2521 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc 2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 2941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 3001 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa 3061 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 3121 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 3181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 3241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 3301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 3361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 3421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 3481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3541 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 3601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 3721 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 3781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 3841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 4021 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 4081 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 4141 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 4201 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4261 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 4321 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 4381 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 4441 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4501 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4561 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 4621 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 4681 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 4741 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 4801 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 4861 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 4921 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 4981 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 5041 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg 5101 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 5161 acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt aattaggctg 5221 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt 5281 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg 5341 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg 5401 aagatcggaa ttccctaggc cggccctgca gagggccctg cgtatg //