LOCUS       sequence-461962-        4860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3288m 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 4860)
  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 4860)
  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: AiP20129 - pAAV-AiE3288m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4860
                     /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      872..881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(878..880,881..883,884..1594)
                     /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(923..944)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1184..1203)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1531..1552
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1687..1707)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1894..2005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2123..2263
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2123..2252
                     /label=AAV2 ITR
     rep_origin      2338..2793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2425..2444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2635..2656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2810..2829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2929..2951
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2989..3007)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3075..3179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3180..3248,3249..4040)
                     /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(3398..3417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4211..4799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4700..4719
                     /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 gcgtttaact gtgttacaag acctttcaaa tctcaaatag
      181 agtgagacaa atagaaccgg ggtgctgggt tcattagcat ttaatgaata acaaattctg
      241 cctgcctggg ctagtaatct gcattagtct ttctcctgag ttttcatttc tgttatcttc
      301 tttcatgcaa attaagaaac tacacaaaca caattagatg tggcagctat ctctgtagca
      361 agttagagga aatgggcttt acaaagcaaa ttacagaata attcttttct tacccgcaat
      421 gatcaaaggc agacattcag gctaatttta gactgtttct gttttcttat aatttagaag
      481 aggaaataat acaaataagt gtgtatgtgt gtgtttgtgt gtgtgtgtgt gtattgtgtg
      541 tgtgtgttta agaaaagaaa tctagctgtt tccgttttct atttttccct catgaatact
      601 tgatttcatg aagtcttaag gaagtggcac accagaggat ttgtgctgaa tccaaggtca
      661 ctgtactctt gggggaatgt acagcgctcc ccaccagcag acaagtaagt tttcttccgg
      721 ctcgcctgca ccatttggct gaatgcgctc agcaacaaaa gctgtgttct cgccggcgga
      781 gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga
      841 tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
      901 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      961 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     1021 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     1081 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1141 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1201 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1261 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1321 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1381 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1441 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1501 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1561 cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     1621 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1681 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1741 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     1801 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1861 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     1921 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1981 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2041 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     2101 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2161 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2221 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2281 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2341 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2401 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2461 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2521 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2581 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2641 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     2701 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2761 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     2821 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2881 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2941 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3001 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3061 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3121 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3181 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3241 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3301 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3361 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3421 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3481 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3541 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3601 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3661 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3721 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3781 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3841 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3901 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3961 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4021 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4081 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4141 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4201 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4261 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4321 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4381 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4441 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4501 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4561 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4621 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4681 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4741 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4801 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
