LOCUS       sequence-432533-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2116m 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 4840)
  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 4840)
  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: AiP1334 - pAAV-AiE2116m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     176..197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(351..370)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     470..492
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(530..548)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        616..720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(721..789,790..1581)
                     /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(939..958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1752..2340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2241..2260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2402..2531
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3253..3262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3259..3261,3262..3264,3265..3975)
                     /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(3304..3325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3565..3584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3912..3933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4068..4088)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4275..4386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4504..4644
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4504..4633
                     /label=AAV2 ITR
     rep_origin      join(4719..4840,1..334)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4806..4825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
       61 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
      121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      181 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
      241 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      301 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
      361 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
      421 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      481 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
      541 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      601 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      661 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      721 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      781 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      841 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      901 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      961 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1021 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1081 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1141 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1201 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1261 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1321 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1381 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1441 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1501 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1561 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2401 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     2461 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     2521 taggggttcc tgcggccgca cgcgttcaca cctggacgac acctgtgaag gtcactgtcc
     2581 ttcactcaca cctggacacc tgtgaaggtt actgtcctcc actcacacct gaatgacacc
     2641 tgtgaaagtc aaggtcacac aaaaaggcac ttttgtgctt atgcttctgt tttaatctgg
     2701 gaacccagat gttctatcag catgctttat tctgaggccc ccgcacatcc ttcatgtatg
     2761 caagccttct cttatgtaac cttttggctt tctgtggcca gatggcttcc tctttcattc
     2821 atccctatgt gaaccaacta tgactccaaa atatctggcc tttgtgtatc acatatactc
     2881 cagctttctg gtctttttta ggttccttga aatatgccag gctgtttcct cttcctgggg
     2941 cttcacagta gcagtctggt atacctggga tattttttct cttatcctca ttatatgtgt
     3001 tcctttctaa atgtcacttc tcatggaatc cactttacct aatttgtgta gattttcaga
     3061 atttatgccc atgccaaaga tctttaccat aactggatct aatcaccttt atgaaggtaa
     3121 gaaaagatcc caaatttatt tagctatttc actgagtttg agctcgggct gggcataaaa
     3181 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
     3241 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3301 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3361 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     3421 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
     3481 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3541 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3601 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3661 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     3721 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     3781 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3841 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     3901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3961 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     4021 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4081 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4141 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     4201 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     4261 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4321 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4381 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4441 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     4501 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4561 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4621 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4681 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4741 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4801 gcccgctcct ttcgctttct tcccttcctt tctcgccacg
//
