LOCUS       sequence-466265-        4671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2184m 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 4671)
  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 4671)
  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: AiP1501 - pAAV-AiE2184m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      118..706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     607..626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   768..897
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1450..1459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1456..1458,1459..1461,1462..2172)
                     /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(1501..1522)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1762..1781)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2109..2130
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2265..2285)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2472..2583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2701..2841
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2701..2830
                     /label=AAV2 ITR
     rep_origin      2916..3371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3003..3022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3213..3234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3388..3407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3507..3529
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3567..3585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3653..3757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3758..3826,3827..4618)
                     /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(3976..3995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
       61 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
      301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
      361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
      421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
      481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
      781 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
      841 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
      901 gccgcacgcg ttgtaagagt aataatcgcc ggcgagggct cactttctct tggttggctg
      961 gccaccgtga ctcagaaatg aaaccgtcac gctccgatgc cccatgcttt tttttcagct
     1021 ttcattttaa taactttatt gacatgtcaa ggtaggcaaa gttgtcaagg gtggtatcaa
     1081 atggtcaaat aaatcaaagc tccatctctc tgcacttccc attacttcag tgtctgtttc
     1141 agacataatc agggtcggtt cacaaatctc tgatgcttta caaatgtcag gcagactttc
     1201 agattatcct tgaagagtct ttctgctagg ttataaaaca ggaacctctc ttatttcata
     1261 cctaggatgg tgttctagtt agagttactg ttgctgtaat gaaacgcaat gactgaaagc
     1321 aagttggaaa ggaaagggtt tattcggccg ccggcggagc tcgggctggg cataaaagtc
     1381 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1441 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1501 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1561 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1621 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1681 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1741 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1801 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1861 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1921 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1981 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2041 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     2101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2161 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     2221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2341 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     2401 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     2461 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2521 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2581 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2641 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     2701 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2761 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2821 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2881 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3121 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3241 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3361 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3421 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3481 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3541 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3601 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3721 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3781 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3841 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3901 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3961 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4021 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4081 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4141 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4201 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4261 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4321 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4381 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4441 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4501 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4561 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4621 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt t
//
