LOCUS       sequence-466231-        5270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2083m 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 5270)
  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 5270)
  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: AiP1273 - pAAV-AiE2083m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..36)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(696..1487,1488..1556))
                     /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     1319..1338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1557..1661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1729..1747
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1785..1807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1907..1926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1943..2398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2080..2101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2292..2311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2473..2613
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2484..2613
                     /label=AAV2 ITR
     polyA_signal    2731..2842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3029..3049
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3142..3852,3853..3855,3856..3858))
                     /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(3184..3205)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3533..3552
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3792..3813
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3855..3864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5016..5145
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(5207..5270,1..525))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
       61 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      361 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1561 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1621 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1681 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     1741 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     1801 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     1861 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     1921 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
     1981 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
     2041 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
     2101 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2161 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     2221 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2281 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
     2341 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     2401 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     2461 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     2521 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     2581 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
     2641 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     2701 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     2761 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     2821 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
     2881 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
     2941 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     3001 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     3061 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
     3121 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3181 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     3241 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     3301 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     3361 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     3421 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     3481 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     3541 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     3601 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     3661 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     3721 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     3781 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     3841 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     3901 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctcgta
     3961 gtcctagtca agagcgctga tcacagagtc cagtagcagt ggcatctatg ggcattccaa
     4021 agctagcagt tggggattcc tgtgtgtgtg cctgtgaaca cattcttgta aatgttccta
     4081 atggaaattc aactctattt tctggtctct taattatctt gcctccagga tttcattttt
     4141 attctcttca ctgtcttgtt ttcccttctc atctcccaca tcccctttgt ggcagtgctg
     4201 gatgctgatg ctgagaaatc tcatgtaacc tccgattcca gctatagaca cgagtaggag
     4261 aaatcaagct taaaaacaaa caaataaaca gaaacagcta aactcaggca tcctgtcaac
     4321 aaaatagaaa ggctagatgg tgaccaagga cactgaaatc catctcagaa gactgggaga
     4381 atgcagtcag atgtggtaaa attccacaga aaatagatat gttgaaaatt ttcactggga
     4441 taaaggtttg gattatcatc atgagtctga ccagctatct ttaacaaaag tggtacacaa
     4501 tttgcatata agcttcattc cttttcccaa ttatgctaaa taaaatgcat tcattttctc
     4561 tgtgattata ggccagacct gccaaacacc tgtgactgcc ataaacccta atttgatatt
     4621 tatttaccca aaccaatctt attatctaaa ttattttagt catcaaaggc cttaattatt
     4681 gagagtagat gcaatagggc tcctgtcctt gggtctttgt ttctgaattg cctctcctgc
     4741 ttagatataa gaggcaatag gattctgcta ctgagggtac tgaaaataac actcttactg
     4801 aatcagaact agatttaaat tataccacag agctgaggtt gataatttca tctggtaacc
     4861 tttagataat tttaaaagtc attatttcta ggccaggtgt ggtggcacat gcctttaatc
     4921 ccagcacttg ggaggcagag gcaggcagat ttctgagttt gaggccagcc tggtctacag
     4981 agtgagttcc aggacagcca gacgcgtgcg gccgcaggaa cccctagtga tggagttggc
     5041 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5101 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag
     5161 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5221 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
//
