LOCUS       sequence-466308-        4672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2537m 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 4672)
  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 4672)
  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: AiP1796 - pAAV-AiE2537m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      230..239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(236..238,239..241,242..952)
                     /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(281..302)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(542..561)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     889..910
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1045..1065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1252..1363
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1481..1621
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1481..1610
                     /label=AAV2 ITR
     rep_origin      1696..2151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1783..1802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1993..2014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2168..2187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2287..2309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2347..2365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2433..2537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2538..2606,2607..3398)
                     /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(2756..2775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3569..4157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4058..4077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4219..4348
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcctccttc ccgcccctcg tggctgcctg agcaggtggc ctttccgctg gaagccaggg
       61 ataaatgagc cagcaggcac aggcccagcc ctgagtgtgg gtcactccag gccaggggtc
      121 gttccccgcc ataccggagc tcgggctggg cataaaagtc agggcagagc catctattgc
      181 ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt
      241 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      301 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      361 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      421 gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
      481 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      541 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      601 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      661 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
      721 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
      781 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
      841 gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
      901 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga
      961 cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga
     1021 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1081 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1141 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1201 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     1261 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1321 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1381 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1441 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     1501 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     1561 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     1621 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     1681 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1741 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     1801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     1921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     2041 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     2101 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     2161 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2221 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2281 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2341 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2401 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2521 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2581 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2641 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2701 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2761 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2821 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2881 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2941 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3001 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3061 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3121 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3181 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3241 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3301 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3361 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3421 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3481 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3541 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3601 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3661 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3721 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3781 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3841 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3901 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3961 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4021 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4081 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4141 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4201 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     4261 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     4321 gtggccaact ccatcactag gggttcctgc ggccgcacgc gttgtaagag taataatcgt
     4381 aatgagagaa gtgtatgctg aatgttctag acacacagtt ttgttcaaat tgccctatga
     4441 taaatgttaa aatctgtgga gaagtgtgag ccatttcaat gaccccaggc cagagagtcc
     4501 cattagcagc ccccaccagc agagttccca gccttccagg ctttgtcgct ccgagatgga
     4561 gagatttctt cccaccagct gtgacctcct tctctccctt gcccccagca caaagccatc
     4621 ccaggacaag tcatggacca aattgccgag cctgtttctc ttggctccca gg
//
