LOCUS       sequence-471216-        4831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2350m 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 4831)
  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 4831)
  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: AiP1766 - pAAV-AiE2350m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4831
                     /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      843..852
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(849..851,852..854,855..1565)
                     /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(894..915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1155..1174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1502..1523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1658..1678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1865..1976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2094..2234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2094..2223
                     /label=AAV2 ITR
     rep_origin      2309..2764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2396..2415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2606..2627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2781..2800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2900..2922
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2960..2978)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3046..3150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3151..3219,3220..4011)
                     /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(3369..3388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4182..4770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4671..4690
                     /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 gcgttgtaag agtaataatc gtgacagaca tgggcaggga
      181 cgggttgtga aagaacaaag gcgtgctggg tgtgccggtg tcaacagttt cctgattcta
      241 acacttaaga caaggcttct aaaaatctag ctggagaaca aaacgactaa ggtgatagcc
      301 aggtaaagtt gtataaaatc ctattctggc aaaaaccttc ctgcagtcac ttgatcaaat
      361 ctgtttttct gatgtaacct gggcccaaac tgggaggttt taaacactga aatcctctct
      421 acataacaag aagtaatttc tcccctcttc ttcaaagaga aatctttcta ttcaatgaaa
      481 ggttttgaat tgaacattca gttatagaat gttcaatttc aagctgttcg gttgaccctc
      541 ggctttctct ccaagttccc ctatgccttt tgcctttaaa agttcagaat gtctccaaca
      601 gtaactgttc aattctgtct tgcaagcctg gactatggac agtgatctaa cctcttggaa
      661 aagacatgga gtgcacccaa aaactgagag agacattctc tcgtcccagc cgccagctca
      721 cgggtttcaa agtcctggga cacacatcgg agctcgggct gggcataaaa gtcagggcag
      781 agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg
      841 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      901 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      961 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1021 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
     1081 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1141 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1201 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1261 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1321 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1381 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1441 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1501 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1561 acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     1621 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1681 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1741 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     1801 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     1861 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     1921 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     1981 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2041 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2101 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2161 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2221 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2281 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     2761 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     2821 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2881 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2941 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3001 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3061 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3121 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3181 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3241 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3301 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3361 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3421 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3481 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3541 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3601 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3661 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3721 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3781 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3841 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3901 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3961 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4021 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4081 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4141 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4201 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4261 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4321 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4381 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4441 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4501 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4561 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4621 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4681 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4741 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4801 ctggcctttt gctggccttt tgctcacatg t
//
