LOCUS       sequence-461977-        5258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2276m 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 5258)
  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 5258)
  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: AiP1609 - pAAV-AiE2276m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1110..1121
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1270..1279
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1276..1278,1279..1281,1282..1992)
                     /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(1321..1342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1582..1601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1929..1950
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2085..2105)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2292..2403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2521..2661
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2521..2650
                     /label=AAV2 ITR
     rep_origin      2736..3191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2823..2842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3033..3054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3208..3227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3327..3349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3387..3405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3473..3577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3578..3646,3647..4438)
                     /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(3796..3815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4609..5197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5098..5117
                     /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 gccggcgtct tctcagactc
      181 cccactttga gctcagatca ggaagccatt ctctaggaac ctgggtttac ttcagctgag
      241 cttgggtcag gtccctgtta aggcctgaga tgccctgcct gcacataaag acatctcacc
      301 atctcacagt ggtcacaacc caggctcctg ccaagcagat tttcaagtct ccaatgcagt
      361 ccctagtcct tctatccact catgcttgaa ggttcctgaa ggaccatagt agggcatggg
      421 aaagcagact ccacagttag cacctcccta acttattgtc cccaactcat caaagctgga
      481 aggctatgaa agagggtagg agttgccctg gagggctgcc tggaggagtg ggggatcagc
      541 ctgcagaagt gactggctgg gtttggttgg agctggctgg ctggctgtgg ggctggctag
      601 cccctctagt ccaggcctgg ccaagcccct agggcagagc tgactcctgg ccattcccgg
      661 gacaacaatg gccagcattt aaattccaat tgagtcggcc agagtggccc cgggcgtgag
      721 aacaggagcc cccaccccca gctccagccc cgcctgccgc attcctgcct ccccggagat
      781 ggccgtggcc aggccgtctc cagggggccc ggaatgcagc actcaatgga agggggacaa
      841 gggcccattg agcagtgacg ggcaggattg agctgtcagc cccccaactc aggacccctc
      901 ctctgtcccc accctctctc cctggtgtta ctgatgacgg ctgctcttgg ggaccccact
      961 catacctcct gtggctgtcc ccgagcgagc atgggcctcg tccatctggc tgactgcagg
     1021 ttctgccctc ttggcccctt tccacccagc agctagggag ctctcgaaac ctcaatcata
     1081 tcatgtcatg cccctgtgta aagcctgtta ttgaaggcag gctagctgac cctgctttcc
     1141 tttgctctgc gtgttctccc ccctcagccg ccggcggagc tcgggctggg cataaaagtc
     1201 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1261 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1321 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1381 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1441 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1501 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1561 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1621 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1681 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1741 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1801 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1861 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     1921 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1981 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     2041 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2101 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2161 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     2221 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     2281 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2341 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2401 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2461 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     2521 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2581 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2641 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2701 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2941 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3061 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3181 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3241 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3301 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3361 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3421 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3781 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4261 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4321 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4501 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4561 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4621 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4681 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4741 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4801 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4861 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4921 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4981 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5041 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5101 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5161 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5221 acggttcctg gccttttgct ggccttttgc tcacatgt
//
