LOCUS       sequence-466283-        4745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2337m 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 4745)
  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 4745)
  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: AiP1656 - pAAV-AiE2337m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      267..276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(273..275,276..278,279..989)
                     /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(318..339)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(579..598)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     926..947
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1082..1102)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1289..1400
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1518..1658
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1518..1647
                     /label=AAV2 ITR
     rep_origin      1733..2188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1820..1839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2030..2051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2205..2224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2324..2346
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2384..2402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2470..2574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2575..2643,2644..3435)
                     /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(2793..2812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3606..4194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4095..4114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4256..4385
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acaggatctg tgtgcagagc cagatcctga gcaaagcaga tgcaatccct gactaaaatg
       61 tttggagaga aagagaactc tctgaaagga tattagctga catttctagt gcctttaaag
      121 tgtacaagat aaagctctct ttctgcaaat atattttttt ccttttatta ccggagctcg
      181 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
      241 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
      301 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      361 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
      421 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
      481 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      541 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      601 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      661 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      721 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
      781 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      841 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
      901 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      961 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     1021 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1081 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1141 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     1201 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1261 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     1321 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1381 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1441 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     1501 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     1561 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     1621 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     1681 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     1741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1801 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1921 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2041 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     2101 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2161 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2221 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2281 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2341 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     2401 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     4261 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     4321 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     4381 ttcctgcggc cgcacgcgtt gtaagagtaa taatcggagt gcaaaaatgt ccatgagata
     4441 ttgaacttgt ttccattgat gtctgttatg ttgcccaaag gagatttttc tttgtagaga
     4501 acaactcatt tctacagaaa cagcactatg agtagatgga agtactcagt gagtcatact
     4561 gaagtgtata atatcacacg gcagacagac caggtcagca ggatcagccg aaggctgaaa
     4621 tatgcatgca ggagatgggt tgcaaacctg agtaagaact cctacatgcc ctcaccctgc
     4681 ttggttcctg aggtacctgc catctcttgc tttacgagca gaatctgtcc ccacttcagt
     4741 attgt
//
