LOCUS       sequence-461965-        4627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3093m 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 4627)
  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 4627)
  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: AiP20050 - pAAV-AiE3093m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4627
                     /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      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(645..647,648..650,651..1361)
                     /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(690..711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(951..970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1298..1319
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1454..1474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1661..1772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1890..2030
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1890..2019
                     /label=AAV2 ITR
     rep_origin      2105..2560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2192..2211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2402..2423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2577..2596)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2696..2718
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2756..2774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2842..2946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2947..3015,3016..3807)
                     /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(3165..3184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3978..4566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4467..4486
                     /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 gcgtctattc caaaatgtaa aattcattta aaaagaaatg
      181 gagaaagacg ccgtgcttcc agaggagact ttgaaaccgg agcctttggt tcccatgaca
      241 gcctcctaac gacactatta tataaaatcc agagaagata ctaaaaacag aatatgcaaa
      301 ttagagaggg gtgggtccgt ggaggagaca ggtaggggag atggcccccc atgcaagggg
      361 gctgtgctgg aggaagcttc cggaacactg agggtggctg gggacagcac ggtgccctca
      421 cgcatgagcc tgactcctgg gccttgtaca gcgtacagcg tgcagctggg gctggactga
      481 aagtcagcag ccgctttacg ggtggaaaga aaaccagaag tgatgaaaga aagaagccgc
      541 cggcggagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      601 tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg
      661 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      721 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      781 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      841 gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      901 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      961 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1021 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1081 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1141 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1201 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1261 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1321 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc
     1381 ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1441 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1501 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     1561 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1621 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     1681 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1741 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     1801 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     1861 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     1921 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     1981 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     2041 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2101 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2161 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2401 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     2461 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     2521 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2581 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2641 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2701 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2761 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3181 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3481 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4621 cacatgt
//
