LOCUS       sequence-470861-        5024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0082h_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 5024)
  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 5024)
  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: AiP14483 -
            pAAV-AiE0082h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5024
                     /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      1036..1045
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1042..1044,1045..1047,1048..1758)
                     /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(1087..1108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1695..1716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1851..1871)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2058..2169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2287..2427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2287..2416
                     /label=AAV2 ITR
     rep_origin      2502..2957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2589..2608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2799..2820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2974..2993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3093..3115
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3153..3171)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3239..3343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3344..3412,3413..4204)
                     /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(3562..3581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4375..4963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4864..4883
                     /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 gcgtgcactg cagctgagca gaatgctgca gtcagggtga
      181 cacaatgcaa actggagcca tgtcctcaag tgcatgccaa tcaaaccaac tttaaccttc
      241 gctggtttgt ggctgctgac ctagagagga atttgttata tggctgaggt cctgtcctac
      301 ctaagccaga ctccagtgca aaaaaaaaat cctgtcagat gcgcctcttc ttctctgtct
      361 ccatctccct ctctgcctcc cgtttcccct ttactcttta ctgttagttg gcactgcagc
      421 tgagcagaat gctgcagtca gggtgacaca atgcaaactg gagccatgtc ctcaagtgca
      481 tgccaatcaa accaacttta accttcgctg gtttgtggct gctgacctag agaggaattt
      541 gttatatggc tgaggtcctg tcctacctaa gccagactcc agtgcaaaaa aaaaatcctg
      601 tcagatgcgc ctcttcttct ctgtctccat ctccctctct gcctcccgtt tcccctttac
      661 tctttactgt tagttggcac tgcagctgag cagaatgctg cagtcagggt gacacaatgc
      721 aaactggagc catgtcctca agtgcatgcc aatcaaacca actttaacct tcgctggttt
      781 gtggctgctg acctagagag gaatttgtta tatggctgag gtcctgtcct acctaagcca
      841 gactccagtg caaaaaaaaa atcctgtcag atgcgcctct tcttctctgt ctccatctcc
      901 ctctctgcct cccgtttccc ctttactctt tactgttagt tggagctcgg gctgggcata
      961 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     1021 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1501 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1561 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1621 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1681 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1741 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     1801 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1861 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1921 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     1981 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2041 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2101 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2161 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2221 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2281 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2341 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2401 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2461 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2521 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2581 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2641 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2701 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2761 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2821 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2881 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2941 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3001 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3061 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3121 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3181 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3421 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3541 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3721 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4021 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4201 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4261 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4321 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4381 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4441 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4501 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4561 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4621 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4681 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4741 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4801 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4861 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4921 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4981 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
