LOCUS       sequence-432532-        4705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2115m 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 4705)
  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 4705)
  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: AiP1333 - pAAV-AiE2115m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      172..181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(178..180,181..183,184..894)
                     /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(223..244)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(484..503)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     831..852
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(987..1007)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1194..1305
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1423..1563
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1423..1552
                     /label=AAV2 ITR
     rep_origin      1638..2093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1725..1744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1935..1956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2110..2129)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2229..2251
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2289..2307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2375..2479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2480..2548,2549..3340)
                     /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(2698..2717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4161..4290
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gacttgtgga gtcttggagc tccaagacat tctgagtctt gagggagact atctcacatc
       61 aactatttca ctgagtttga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      121 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      181 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      241 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      301 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      361 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      421 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      481 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      541 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      601 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
      661 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      721 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      781 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      841 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc
      901 gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
      961 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1021 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1081 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1141 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     1201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1381 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     1441 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1501 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     1561 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     1621 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1681 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     1741 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1801 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     1861 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1921 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     1981 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     2041 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2101 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2161 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2221 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2281 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2341 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     4201 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     4261 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtataata tattaatgaa
     4321 catgataaag taataattat atgattaata tattagaggg aatgtaatag agcgaagctg
     4381 cttctgtgcg tcctttgagc gtctgcacct gggcctgaga gagactccga gcagctgcct
     4441 gtgcagcagt gtgagcctgc caggttgtgc ttggggacac cagtgacacc agctgctcct
     4501 catatctcca cacgaggcta ttttaagaaa ctccgcactt gatcagggag catgtggggt
     4561 taagccttat gttttccact ttggttttta gatcctaagc aggtgctgca gttaaaagtt
     4621 caaaattttt tttaaaaagt cttcttttta accattaaga ccattaagat aagttcctgt
     4681 cggaggtagc attaagtaaa gactg
//
