LOCUS       sequence-461945-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2069m_3xC2 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 5150)
  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 5150)
  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: AiP1328 - pAAV-AiE2069m_3xC2-minBG-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5150
                     /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      1162..1171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1168..1170,1171..1173,1174..1884)
                     /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(1213..1234)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1474..1493)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1821..1842
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1977..1997)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2184..2295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2413..2553
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2413..2542
                     /label=AAV2 ITR
     rep_origin      2628..3083
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2715..2734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2925..2946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3100..3119)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3219..3241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3279..3297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3365..3469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3470..3538,3539..4330)
                     /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(3688..3707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4501..5089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4990..5009
                     /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 gcgttcctta ggccatccct caccaaagag cctatttctg
      181 tcttctgtcc atgccaaata aatgattctg agtggtttgc cttgagtcca caggagaggt
      241 atgcttttag ttcatatata agaaatgctc gttgtttttt gtgccatggg aaatgcaaat
      301 gtttccccga tggagaaatg gcccatgttc ttggccaaat cattgagatc tctgactgga
      361 atgataagtg agcaaacatg tggtgtctcc cctactccaa tctacctttg ctttatcttt
      421 tagattggct tgggcaaaag aatccagaga tccttaggcc atccctcacc aaagagccta
      481 tttctgtctt ctgtccatgc caaataaatg attctgagtg gtttgccttg agtccacagg
      541 agaggtatgc ttttagttca tatataagaa atgctcgttg ttttttgtgc catgggaaat
      601 gcaaatgttt ccccgatgga gaaatggccc atgttcttgg ccaaatcatt gagatctctg
      661 actggaatga taagtgagca aacatgtggt gtctccccta ctccaatcta cctttgcttt
      721 atcttttaga ttggcttggg caaaagaatc cagagatcct taggccatcc ctcaccaaag
      781 agcctatttc tgtcttctgt ccatgccaaa taaatgattc tgagtggttt gccttgagtc
      841 cacaggagag gtatgctttt agttcatata taagaaatgc tcgttgtttt ttgtgccatg
      901 ggaaatgcaa atgtttcccc gatggagaaa tggcccatgt tcttggccaa atcattgaga
      961 tctctgactg gaatgataag tgagcaaaca tgtggtgtct cccctactcc aatctacctt
     1021 tgctttatct tttagattgg cttgggcaaa agaatccaga gacttaagga gctcgggctg
     1081 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     1141 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1201 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1261 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1321 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1381 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1441 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1501 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1561 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1621 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1681 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1741 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     1801 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1861 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     1921 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1981 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2041 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2101 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2161 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2221 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2281 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2341 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2401 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2461 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2521 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2581 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2701 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2761 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2821 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2941 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3001 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3061 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
