LOCUS       sequence-432566-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2555m 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 4739)
  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 4739)
  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: AiP1846 - pAAV-AiE2555m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      573..582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(579..581,582..584,585..1295)
                     /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(624..645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(885..904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1232..1253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1388..1408)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1595..1706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1824..1964
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1824..1953
                     /label=AAV2 ITR
     rep_origin      2039..2494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2126..2145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2336..2357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2511..2530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2630..2652
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2690..2708)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2776..2880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2881..2949,2950..3741)
                     /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(3099..3118)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3912..4500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4401..4420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4562..4691
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cagatggact ttcttttgca tctgctgtgt acaggctggg ctttgtactc ttttcctggg
       61 ggagagaaag tctttcttct ctgctagttg attcctgttt ttgtgaagca gctaagaaaa
      121 catctgtggc aggcagaaaa aattgacaaa ggattgacaa acaccattag ctgaaaattt
      181 tccatttcat tgcaactatg cattgtttac tttactgtaa atatcttaat acatcatcct
      241 ctgaatatgc tggcagagag gctggagaac cgtgatttca attaaggtta gtaattgatg
      301 gtatctagcg ttcaaagact ggggcttgat taacagctgc ttggacaaat tgtcattgtg
      361 aagtggtttt gatgtgcatt gaagattatt tctttctggc ttatctgatg ttgtggctgt
      421 gaaatgcttg tcttggattt gcttattttt gtaaactact tcctttggct gtaaattcgg
      481 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
      541 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      601 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      661 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      721 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      781 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      841 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      901 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      961 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1021 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1081 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1141 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1201 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1261 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1321 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1381 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1441 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     1501 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1561 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     1621 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1681 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1741 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     1801 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     1861 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     1921 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     1981 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2101 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2221 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2341 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2401 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2461 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     2521 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2581 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2641 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2701 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2821 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2881 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2941 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3001 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3061 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3121 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3181 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3241 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3301 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3361 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3421 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3481 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3541 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3601 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3661 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3721 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3781 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3841 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4021 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4081 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4141 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4201 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4261 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4321 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4381 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4441 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4501 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4561 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     4621 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     4681 taggggttcc tgcggccgca cgcgttgtaa gagtaataat cggtgttgtg agctgctag
//
