LOCUS       sequence-466344-        4790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2635m 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 4790)
  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 4790)
  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: AiP2103 - pAAV-AiE2635m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(291..310)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     410..432
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(470..488)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        556..660
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(661..729,730..1521)
                     /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(879..898)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1692..2280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2181..2200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2342..2471
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3143..3152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3149..3151,3152..3154,3155..3865)
                     /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(3194..3215)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3455..3474)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3802..3823
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3958..3978)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4165..4276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4394..4534
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4394..4523
                     /label=AAV2 ITR
     rep_origin      join(4609..4790,1..274)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4696..4715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
       61 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      121 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
      181 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      241 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
      301 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
      361 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      421 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
      481 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      541 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      601 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      661 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      721 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      781 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      841 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      901 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      961 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1021 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1081 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1141 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1201 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1261 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1321 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1381 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1441 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1501 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1561 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1621 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1681 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1741 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1801 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1861 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1921 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1981 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2041 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2101 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2161 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2221 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2281 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2341 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     2401 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     2461 taggggttcc tgcggccgca cgcgtattag tttttacaac cttttgtaat ttggtattgt
     2521 atacaacatg gtcatatgcg gagtttctgt aatcttttca gtgggtgata gaaatatgag
     2581 aagcaaattt ataagccatt tagcgaagca tttaatcaat ttcatgcata ggaaacaact
     2641 ctgatgcctt ggctttatca atctttctct ccagcagaac cactaaccca ctcggtctcc
     2701 aaacacagcc tgggtggggg aagggtggag accaaggagg aaccttttcc tgtggcagaa
     2761 tggaaatgtc agggcttctg aagccggcct gacgcactta gcagcgtctc aaaaccttct
     2821 ggaggctgtt aacgtatttt agacaaggag ctgggatctg gtgttcggag gaagttttca
     2881 aggcctttta ctgcaaagtt attcattaaa caccctttac cagagtgact ctcacgtgct
     2941 cagccataca gattggactc agctttgggc tccaagaaat atttttaaca aatccattgc
     3001 tttcaaatgc atggaaaaga agaaacaaaa caaaactatt tcgccggcgg agctcgggct
     3061 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     3121 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3181 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3241 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     3301 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     3361 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3421 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3481 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3541 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3601 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     3661 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3721 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     3781 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3841 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     3901 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3961 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4021 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     4081 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4141 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4201 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4261 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4321 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     4381 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4441 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4501 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4561 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4621 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4681 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4741 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
//
