LOCUS       sequence-425377-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2443m 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 5126)
  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 5126)
  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: AiP1869 - pAAV-AiE2443m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   609..738
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1746..1755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1752..1754,1755..1757,1758..2468)
                     /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(1797..1818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2058..2077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2405..2426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2561..2581)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2768..2879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2997..3137
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2997..3126
                     /label=AAV2 ITR
     rep_origin      3212..3667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3299..3318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3509..3530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3684..3703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3803..3825
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3863..3881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3949..4053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4054..4122,4123..4914)
                     /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(4272..4291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5085..5126,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
      661 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      721 ccatcactag gggttcctgc ggccgcacgc gttgtaagag taataatcga ctggccaatg
      781 gtctcacctg ttctacctcc ccatggctgg gtttccagtt gtgcacggga actcaagact
      841 tttacataag tcctagggat gcaacctagt gtcctcatgc ttccatggca ttccctttaa
      901 gaactaagct atctccacag ccctgctttg agttgtcaat ctttcattct ctgcaaagaa
      961 tctattagat gagttcagct ttgtcctggt tagaacaaaa ggaagtatgc cttgtttgtg
     1021 atcctgcaga atggagaggg ttgggaaact ccatatactt tactgacaaa taaagcaaac
     1081 atgaaacact gaaatcccat atgttgtgag gcacgctggc agtctgcctc atccccaacc
     1141 cagtgcaagc aggctggttt gcaatctgga gcctccctcg ctgagctgta acaaaattga
     1201 tcctaggtgc atgtaagaca cagcgcattc cttcccgtgc tgatttatcc ctggcctctt
     1261 ccctacattt ctccttttag cccatcattt agtctctgat ataactcaca ggcaaccatt
     1321 gaaaataatg cagggggtta cttctcagca gcagcagtgg tggagatcct tctgcctgta
     1381 tgagcaaaga ggccatgtgg agccagcaag ccttcccaac gaaggtcagg tgtaaagcgt
     1441 agattaccaa cagcactagg aagacaagca ctgcattgat gtcattcacc ctaatagccc
     1501 caccccctgg caccatgcct ggaatgagag ggatactaaa aaaatcattt atggaatgac
     1561 tcaagaaaag gcaaaggagt tggctcatcc agtaaagtgc ttacctagca aacctgagaa
     1621 cctgagctca gtcccaagaa cccatatcaa cggagctcgg gctgggcata aaagtcaggg
     1681 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1741 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1801 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1861 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1921 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1981 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2041 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2101 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2161 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2221 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2281 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2341 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2401 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2461 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     2521 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2581 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2641 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     2701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     2761 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2821 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2881 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2941 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     3001 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3061 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3121 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3181 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3241 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3301 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3361 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3421 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3481 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3541 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3601 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3661 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3781 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3841 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3901 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4021 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4081 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4141 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4201 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4261 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4321 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4381 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4441 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4501 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4561 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4621 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4681 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4741 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4801 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4861 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4921 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4981 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5041 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5101 ctgcgcgtaa tctgctgctt gcaaac
//
