LOCUS       sequence-470304-        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2032m 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 5108)
  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 5108)
  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: AiP1274 - pAAV-AiE2032m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1120..1129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1126..1128,1129..1131,1132..1842)
                     /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(1171..1192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1432..1451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1779..1800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1935..1955)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2142..2253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2371..2511
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2371..2500
                     /label=AAV2 ITR
     rep_origin      2586..3041
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2883..2904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3058..3077)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3177..3199
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3237..3255)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3323..3427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3428..3496,3497..4288)
                     /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(3646..3665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4459..5047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4948..4967
                     /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 gcgttgctgt catttaccta cggttgtctc caaatttctt
      181 caaccaagta gagaaaaatg agagagaagg aaagaaaaaa agaggtatgg ggagaagaga
      241 aagaaggcaa cttgttaaaa atctcagtca aacttacata ctatatagaa cagcatggtg
      301 aatttagggc acatggatat aaaatggaag tttcttattc agtagcagca acttgtgggc
      361 acaggagttg gcaaagataa aaatgtccaa agtcacaaat acaatgtata gttagtcata
      421 ggtgctgtta tttgcctcaa aaaatagact tttattttgc ctttcttttc tttaaccaca
      481 ctcaaaatta gagaacagag acaaaaccca gcaggaaata gcacagaaag cccacagaat
      541 caaagacgtg ttcaaacagc cagctgaatt cattgcacat ttcaaccaca gaaatatttt
      601 caggtgattc tgttgtttga caaaacgtgg gaaccacagg atctacaaca cttgcaagca
      661 aaactcaaca gctctaataa tagttacaga agtgaaagcc aatttggata aaataagaca
      721 ttgactcaag tcctctcaga agagttttga aagcaaagtt tacaaaagtc tggtttgtcc
      781 tttgggattt acagaccttt cagccccttg attcattttt tttttttgga tttcttcatc
      841 actgggagaa ttcccatgca ttatttctcc cctgcttcaa aatcatcaaa tgtgaaacat
      901 ttttcactct tttcttctgt atatagtgat aaaatagcta ttggcttttg gctaaatgtg
      961 ctactttgag cccacccaca caagggagaa atgggggcag acatgagttt gggcatgagt
     1021 cttaaggagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
     1081 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
     1141 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1201 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1261 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1321 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1381 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1441 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1501 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1561 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1621 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1681 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1741 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1801 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg
     1861 cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact
     1921 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1981 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     2041 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2101 ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca
     2161 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2221 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2281 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2341 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2401 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2461 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     2521 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     2581 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2641 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     2701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2881 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     2941 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3001 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3061 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3121 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3181 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3241 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3421 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3481 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3541 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3601 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3661 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3721 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3781 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3841 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3901 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3961 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4021 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4081 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4141 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4201 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4261 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4321 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4381 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4441 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4501 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4561 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4621 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4681 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4741 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4801 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4861 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4921 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4981 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5041 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5101 tcacatgt
//
