LOCUS       sequence-470132-        4761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2465m 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 4761)
  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 4761)
  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: AiP1775 - pAAV-AiE2465m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4761
                     /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      773..782
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(779..781,782..784,785..1495)
                     /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(824..845)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1085..1104)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1432..1453
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1588..1608)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1795..1906
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2024..2164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2024..2153
                     /label=AAV2 ITR
     rep_origin      2239..2694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2536..2557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2711..2730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2830..2852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2890..2908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2976..3080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3081..3149,3150..3941)
                     /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(3299..3318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4112..4700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4601..4620
                     /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 gaaacgagga attatactct
      181 aagcacaaca caagttttct gcatttgttc ctgaagaaag tccttatttt ctaaggtact
      241 cctaggccct gtctataaca ttataatcat cttctagttc atcacagcta agtaaagatt
      301 aaagcattgc acatccagct aagtactttt ctttttttaa aaaagagttc atttcctctc
      361 tcttgaactt gtttgctata aatgtatata aatcaggcat gatgacaatt taataagaac
      421 ttcaagacct gttcatccca tgggatcact actgtgacgt ctttcgggga gccttctgat
      481 tgctcagccc gaagcacaag ccctataaaa tgagagactt tgagatgctg gtcagcttgg
      541 acagcagagc actagccagc tacagccaac cgttccccag gaaccgggaa cagactggtg
      601 gagccttccc tagagcagaa cttcagcacc ctagacagct gccacgaagc cggaaaggca
      661 gccctgcctg aaggaaacgg agctcgggct gggcataaaa gtcagggcag agccatctat
      721 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
      781 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      841 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      901 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      961 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1021 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1081 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1141 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1201 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1261 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1321 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1381 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1441 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt
     1501 cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg
     1561 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1621 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1681 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1741 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     1801 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1861 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1921 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1981 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     2041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2161 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2281 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     2581 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     2641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     2701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     2761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3781 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4741 gctggccttt tgctcacatg t
//
