LOCUS       sequence-466246-        5741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2136m 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 5741)
  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 5741)
  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: AiP1365 - pAAV-AiE2136m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1426..1435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1432..1434,1435..1437,1438..2148)
                     /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(1477..1498)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1738..1757)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2085..2106
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2241..2261)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2448..2559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2677..2817
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2677..2806
                     /label=AAV2 ITR
     rep_origin      2892..3347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2979..2998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3189..3210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3364..3383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3483..3505
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3543..3561)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3629..3733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3734..3802,3803..4594)
                     /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(3952..3971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4765..5353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5254..5273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5415..5544
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctctacgatg agtagctaag ggagaaaata tgcttgctgg caggatcccc ttggggataa
       61 ctgtaggccg gtcctatttc ataaaatagc acacagaagt aggagactat cggtgagtgg
      121 cagggaaagc aggcttcaca gagatgttga gtaactacag gatgtttagt ctagagaaga
      181 aagattatgg ggactttgat ggttgttttc atcatttcaa gaactattgt gtgactacaa
      241 ggagaatctt gagacctact ggggctattc cagttggcca atagagtgag aatgttagag
      301 agagattcgt tttgatgtct tcggaagcct gctctaaagg tatctctgcc tgtgagcatt
      361 ccagaagctg gctgaccgcc cggcctataa agaaacaagt ttttgccagt aattccagtg
      421 aatgatcttt gaggactcct cagacgttac tccttttgtt cttgtcctcc aaaagaaatg
      481 gaaaataaca acactttccc tgggtcttcc tgacgctttc agagtgacaa gaatgttacc
      541 acagggcact gtgtgttccc accttccccc accttcaatg gccggccact gcagcaacct
      601 tgacttagtt aactgaatct gttacagtca acagaccttg gtcccagtga cattcttcct
      661 gccctggtca catactgctg agcctgtgaa ccgttactaa gacctcaact gtgattgctt
      721 tggcccaccc actggacata caaattctag tgctgaacag aacagtcaag aaagaggggg
      781 tatcttgcct caaggtgagg taaatggtag gaagacgttc acagagcagt gcccacgaga
      841 ggcagtgcac acaaaggttc cacgtacagc ccacagcttg acgtaggagt gtccattctc
      901 tgagggaatg tcaaaggaag ggaggtgcct ctcagtgccc tgcagctgcc ctaatcttga
      961 ctgtggctgc atctcccagg ctttgcagtc tagactacca tctcctgaac acctgctttg
     1021 ttttttggtt ttgttttttt tttcgtaaaa ctttcccaga cgtcatcaca caacttggaa
     1081 aagagcgtgt ctcctgagtt gccatttctg ttgtcacgtt ctgaccagcc aacgcttgga
     1141 aaacaaacat acagttcccg ttggaaagac tgtggtaata tttcttcaca gaagtttgct
     1201 acctcaggaa accagttacc tctttgtttc ttgctggata catggttttg gcaccatgtc
     1261 aaacctgcct gaggaatcac tgtttacctc agaggatgta gccttgctgc tttagcagtg
     1321 tggcaggctt aggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1381 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
     1441 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1501 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1561 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1621 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1681 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1741 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1801 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1861 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1921 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1981 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2041 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2101 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     2161 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     2221 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2281 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2341 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2401 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     2461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2641 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2701 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2761 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2821 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2881 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2941 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3241 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3301 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3361 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3421 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3481 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3541 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3601 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5401 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     5461 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     5521 ccaactccat cactaggggt tcctgcggcc gcacgcgttt tcaatagctt gaatccagga
     5581 tggagggggg aaaaaagaaa cctgtaattt tcacattcaa atacaatcct gcggactttt
     5641 aattaaccta actgttccta caatgcttgc tgagtaccat gcacctgcag gccagcactg
     5701 tggccttgcc tcctgacgca cctgcagatc tcatcctgat g
//
