LOCUS       sequence-466273-        5097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2274m 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 5097)
  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 5097)
  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: AiP1607 - pAAV-AiE2274m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      443..452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(449..451,452..454,455..1165)
                     /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(494..515)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(755..774)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1102..1123
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1258..1278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1465..1576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1694..1834
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1694..1823
                     /label=AAV2 ITR
     rep_origin      1909..2364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1996..2015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2206..2227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2381..2400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2500..2522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2560..2578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2646..2750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2751..2819,2820..3611)
                     /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(2969..2988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3782..4370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4271..4290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4432..4561
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 atggacccat cgatgtgggt caagagaacg atcttaggta agacagggtg gagcgagggc
       61 cgtccggggg tgagaaagtc cagcacactc aagtttgtgc gcagccctca ggtgtcagcc
      121 aagatgcggg ggcgtgtgca gcgactccag tgcgtccata gcaccaatgc ggcagattcc
      181 tgagtaccct gaaccataac accctgaacc aactatggtt cagggagctg ggatgggagg
      241 acagttcagc tggaggatat tctgtcgttt ggctaccccc agggacgccc ctgagaagtt
      301 ttcctgcaga cctaggaaac cttcatagat gtagataccg ccgccggcgg agctcgggct
      361 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      421 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      481 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      541 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      601 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
      661 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      721 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      781 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      841 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
      901 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
      961 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1021 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1081 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1141 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1201 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1261 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1321 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     1381 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1441 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1501 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1561 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1621 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     1681 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     1741 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     1801 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     1861 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     1921 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1981 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2041 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2101 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2161 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2221 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2281 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2341 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2401 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2461 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2521 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     2581 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2761 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2821 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2881 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2941 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3001 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3061 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3121 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3181 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3241 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3301 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3361 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3421 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3481 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3541 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3601 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3661 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3721 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3781 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3841 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3901 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3961 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4021 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4081 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4141 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4201 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4261 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4321 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4381 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     4441 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     4501 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     4561 tgcggccgca cgcgttgtaa gagtaataat cgccggcggt cacaggcacc cccagtaacc
     4621 tttccaaacc tatctgccct ccctgcatgc cctggagcag gaaggtacag gtctgaccct
     4681 tataaaaagc tgaggcactg caaagcgaag tctgtcgcac agaactaggg acagatccgg
     4741 attctctggg cttccccgtg gggcctccca gggcacctgc ccaaggccac ccagagcgcc
     4801 agcccaggca gcgccggtgg agcggagccg gacctcccag cggccgatgg gggcggtgac
     4861 cgcccggggc cttcccacgg ttcggtggcg cgtcaccgcg ggggcgaggg ggcgccgggg
     4921 cgccccgggc tgcggtgcag gaaagggccc tgggaaaggg cggcgtcggc tcccgggccg
     4981 gggcggggct gaggcggggc ccaacaacca actccaggga tcgcagggct gggaccccgc
     5041 agccagcttt cggcatgaag cggcccaggg cgcccagtgg ctccgacggc gagtctg
//
