LOCUS       sequence-463075-        4440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3092m 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 4440)
  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 4440)
  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: AiP20177 - pAAV-AiE3092m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4440
                     /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      452..461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(458..460,461..463,464..1174)
                     /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(503..524)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(764..783)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1111..1132
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1267..1287)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1474..1585
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1703..1843
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1703..1832
                     /label=AAV2 ITR
     rep_origin      1918..2373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2005..2024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2215..2236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2390..2409)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2509..2531
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2569..2587)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2655..2759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2760..2828,2829..3620)
                     /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(2978..2997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3791..4379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4280..4299
                     /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 gcgtaacagc tggcctccat ccctgcttct cagtcttgcc
      181 agtcatagtt cctatccagg gcctttggca ccaccttggg acagtggcat cttctaggca
      241 ctgtttcctc ccggaaactt cttcccttca gtgagataaa agagtctgac agaaatctct
      301 ccaggaacac aaggctttct ctcaggtgag gagacaccat taaagctaaa cgccggcgga
      361 gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga
      421 tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
      481 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      541 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
      601 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
      661 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      721 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      781 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      841 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
      901 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
      961 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1021 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1081 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1141 cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     1201 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1261 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1321 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     1381 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1441 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     1501 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1561 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1621 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     1681 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     1741 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     1801 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     1861 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     1921 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1981 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2041 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2101 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2161 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2221 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     2281 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2341 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     2401 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2461 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2521 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2581 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2941 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3001 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3061 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3301 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
