LOCUS       sequence-461994-        4634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2162m 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 4634)
  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 4634)
  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: AiP1436 - pAAV-AiE2162m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4634
                     /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      646..655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(652..654,655..657,658..1368)
                     /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(697..718)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(958..977)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1305..1326
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1461..1481)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1668..1779
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1897..2037
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1897..2026
                     /label=AAV2 ITR
     rep_origin      2112..2567
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2199..2218)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2409..2430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2584..2603)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2703..2725
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2763..2781)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2849..2953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2954..3022,3023..3814)
                     /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(3172..3191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3985..4573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4474..4493
                     /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 gccggcggag aagaatggca
      181 agtgtgtgag agagggcaaa gcagaaaagg gccaagcggg tcctcaggga gaagccacag
      241 ccctctccag cctgtgctgt ccaggaacaa agctgcccta ggcctcccca ggggctcact
      301 ggccggtctc tgggtttccc cggctaagag ctcccggcac aggggaggag ccgggagtgg
      361 gcggggtcgt gaacacgggc tggggcgggg ctagaggggg agggctcagc cttagtccct
      421 ttgggtgtct agagggcggg gcctaggctg atggagatcc gtgaccctag aggatacact
      481 ctgaaggagg tcagggccac gacagagaag gggaggaggc aggcggctgt gcaaataaag
      541 gtttcgccgg cggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
      601 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
      661 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      721 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      781 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      841 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      901 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      961 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1021 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1081 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1141 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     1201 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1261 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1321 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     1381 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     1441 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1501 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1561 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1621 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     1681 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1741 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1801 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1861 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     1921 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     1981 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2041 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2101 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2161 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2221 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2281 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2341 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2401 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     2461 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     2521 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     2581 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2641 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2701 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2761 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2821 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2941 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3061 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3121 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3241 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3361 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3481 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3541 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3661 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3781 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3841 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3901 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3961 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4021 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4081 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4141 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4201 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4261 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4321 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4381 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4441 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4501 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4561 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4621 ttttgctcac atgt
//
