LOCUS       sequence-470466-        4942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0483m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4942)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4942)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP12431 - pAAV-AiE0483m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      137..725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     626..645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   787..916
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1740..1749
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1746..1748,1749..1751,1752..2462)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1791..1812)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2052..2071)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2399..2420
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2555..2575)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2762..2873
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2991..3131
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2991..3120
                     /label=AAV2 ITR
     rep_origin      3206..3661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3678..3697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3797..3819
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3857..3875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3943..4047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4048..4116,4117..4908)
                     /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(4266..4285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
       61 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      781 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
      841 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      901 atcactaggg gttcctgcgg ccgcacgcgt gtggaggttg ggtgtttcca tcacctggct
      961 aacagagatc acaatctcct tttgttcatc ttagggataa ggtttgtcca tagctcgact
     1021 tgtctggatt tctctgtcca tcacatgcct atgtacgctc cccacacctc tctcctagtg
     1081 gctccttggt ttaaaatgcc ccagatctgc agctcacaga agaacatttg gctactcccg
     1141 tttttaatct actacttctg taatggggct cgggagctgg gctgggtccc ccgaacctgt
     1201 gggcagctgg cagcctgacc ctgggctgag taccaggcat ggggaggcca gaaggaaggg
     1261 gcagattgag ccagagaggt aagactgcag agagccaaat aggaggggag gaggccagag
     1321 cctgagggtc cagatggggc tggctgggca caggagggca tctgatgcaa tccgccagga
     1381 gagatgccag ccggccgggt gctgcagagg ctctgttctt gaagagtcta gtgcctgctc
     1441 tattaattct ggctctgtgg gaattgggga agatttccct atgaggtctc ctgagcccca
     1501 gcaacctccc caaaggcctt agctcataaa agcacctgtg tctgaacaag cagggggtgg
     1561 ggacatagaa cacctgggtt ggtggctgcc tggggaaggg caagctgaag tatcttgtca
     1621 gtataaacac agtacctcag ccctgcgagc tcgggctggg cataaaagtc agggcagagc
     1681 catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg
     1741 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1801 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1861 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     1921 ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca
     1981 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2041 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2101 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2161 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     2221 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     2281 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2341 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     2401 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2461 agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     2521 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2581 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2641 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     2701 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     2761 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2821 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2881 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2941 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     3001 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3061 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3121 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3181 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3541 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3661 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3721 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3781 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3841 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3901 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3961 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4021 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4081 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4141 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4201 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4261 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4321 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4381 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4441 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4501 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4561 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4621 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4681 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4741 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4801 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4861 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4921 gtttactcat atatacttta ga
//
