LOCUS       sequence-461946-        5050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2124m 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 5050)
  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 5050)
  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: AiP1353 - pAAV-AiE2124m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5050
                     /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      1062..1071
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1068..1070,1071..1073,1074..1784)
                     /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(1113..1134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1374..1393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1721..1742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1877..1897)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2084..2195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2313..2453
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2313..2442
                     /label=AAV2 ITR
     rep_origin      2528..2983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2615..2634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2825..2846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3000..3019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3119..3141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3179..3197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3265..3369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3370..3438,3439..4230)
                     /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(3588..3607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4401..4989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4890..4909
                     /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 gcgtgtagaa aatggctgga gttcggatgc aggcaccctg
      181 gctgattcag acactggaca ggaagcacta ctgcctgccg tttcttggga agcgacttcc
      241 agtctccaaa cattttcctc tgctgtgcaa tgggcatcca cacacttagc ctcctgtcct
      301 gccccgtagg gttgagcatc ctcagacttg gaccctgtgg gatctgcagc caggtgcaga
      361 ggctgtgcac acctgtaact tcagtacttg gggtgtggag acaagaggag caggagctga
      421 aggtcatcct tggctaaaca gagttcaaag tcatcctaag ctatgtgaga cactcataca
      481 ggaatacatt cttaaaagga acttttgtta gccccacctg caggaagtaa gtttgtgcca
      541 aagccctggg ccccttacct atgcaagttc ctgttaggtt acaccatccc ccactgagat
      601 ctaagccttg gaaaccccag gaaacaactg aatgtccagt cctttgtgcc cttcctgatg
      661 agagagcctc tctccccacc tactgacact tagcaaaggc tccatcacgg ccccacctct
      721 ggttaaagga ggaagctggg tctgggtcgg aaagtcccgc ccaggtgtga agcaagccct
      781 tcctctgccc acacccttca ccatgcccag aagataaagt acacaggatg agggccagct
      841 catacacagc cctgcagtcc tgtgcggcac gtccagcatg ttgtttgcct gtgctctcct
      901 tgccctcctg ggtctggcaa cctcctgcag tttcatcgtg ccccgcagtg agtggagggc
      961 cctgccatga gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     1021 gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg
     1081 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1141 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1201 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1261 tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1321 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1381 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1441 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1501 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1561 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1621 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1681 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1741 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc
     1801 cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga
     1861 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1921 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1981 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2041 ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc
     2101 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2161 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2221 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2281 catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     2341 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2401 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     2461 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     2521 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2581 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     2641 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2701 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     2761 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2821 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     2881 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     2941 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3001 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3061 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3121 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3181 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3541 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3781 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4201 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4261 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4321 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4381 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4441 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4501 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4561 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4621 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4681 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4741 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4801 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4861 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4921 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4981 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5041 gctcacatgt
//
