LOCUS       sequence-470426-        5244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2137m 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 5244)
  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 5244)
  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: AiP1366 - pAAV-AiE2137m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5244
                     /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      1256..1265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1262..1264,1265..1267,1268..1978)
                     /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(1307..1328)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1568..1587)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1915..1936
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2071..2091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2278..2389
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2507..2647
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2507..2636
                     /label=AAV2 ITR
     rep_origin      2722..3177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2809..2828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3019..3040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3194..3213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3313..3335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3373..3391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3459..3563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3564..3632,3633..4424)
                     /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(3782..3801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4595..5183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5084..5103
                     /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 gccggcgcac ctcattaact
      181 cccaggcact aagcttagat cacctttctg tggtgacctt tttagctccc ttccctttct
      241 ccaagttatc ttccagggga aggagctagg cctggtctta ggggacagga gtgagcccct
      301 tggggaccag ggctgggccc tttggctgcc atctggcagt acagtctgga gcctcccaag
      361 cccctgctgt gaggcctctg gggtgaaagt gtctgtcagg ctgtgaggtg ggtctgtgcc
      421 cagttcactc tgactatcca gagacttggg ctgttggcct ggctcggaga acgggttctg
      481 ctgccctggg cacaatgcag agcaaagagg aaagaaacct tttccaaagc agaggccagg
      541 aggaagcaca cagaggcctc tttgctttgg aaccgaagcc tttcaggtct gttttcttta
      601 aatatcctgc tacaggaaag agaagcccaa atccagacat tcctaaaaca ttcaggtggc
      661 tttaaagcag cagtttgttt gatcttcacc ccaccctgtt tctgctcact ggggtctctg
      721 ggacctactc ctgcctggag cttggttgga aggtccacac ccttccaagt tcactcacat
      781 agccgatgta ccgatggggc ctgagacctg attggaactt gctttttcta ctttagtctg
      841 tttttgtttt cgtctgccgt gctgaagatc aaactcttgg ccgtgtgcat cctaggcgct
      901 cactaccact gagtctcagc ccctgcccag gttttgtttg gtttggtttg aagtcacttt
      961 tctttgatcc ccagctgtaa tggcatggct gaccacaccc ttttcctgga aagtctttcc
     1021 cttgacctct ggggtcagac acacaatgac ttctgactca gtgctctgcc tcactactga
     1081 tcactagacc taaagtccca tcatgaggca cctgagggtc tccatcactt cactggtctt
     1141 taacccagag gactcgccgg cggagctcgg gctgggcata aaagtcaggg cagagccatc
     1201 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
     1261 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1321 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1381 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     1441 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1501 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1561 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1621 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1681 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1741 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1801 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1861 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1921 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1981 agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat
     2041 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2101 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2161 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2221 ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg
     2281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2461 aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg
     2521 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2581 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     2641 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2701 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2761 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     2821 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2881 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     2941 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3001 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3061 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     3121 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     3181 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3361 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3421 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3541 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3601 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3661 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3721 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3781 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3841 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3901 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3961 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4021 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4081 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4141 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4201 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4261 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4321 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4381 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4441 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4501 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4561 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4681 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4741 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4801 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4861 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4921 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4981 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5041 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5101 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5161 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5221 tttgctggcc ttttgctcac atgt
//
