LOCUS       sequence-470597-        4523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0539h 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 4523)
  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 4523)
  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: AiP13192 - pAAV-AiE0539h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..159
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(197..219)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     319..338
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(355..810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(492..513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     704..723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   885..1025
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   896..1025
                     /label=AAV2 ITR
     polyA_signal    1143..1254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1441..1461
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1554..2264,2265..2267,2268..2270))
                     /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(1596..1617)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1945..1964
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2204..2225
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2267..2276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2681..2810
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2872..3460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2952..2971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3631..4422,4423..4491))
                     /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     4254..4273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4492..4523,1..73))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
       61 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
      121 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
      181 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
      241 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
      301 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
      361 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
      421 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
      481 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
      541 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
      601 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
      661 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
      721 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
      781 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
      841 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
      901 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      961 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     1021 ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct tcccaatcct
     1081 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     1141 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
     1201 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgaag
     1261 atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca
     1321 aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata caaggaggag
     1381 aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca
     1441 catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa
     1501 tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac
     1561 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     1621 tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg
     1681 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
     1741 acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc
     1801 ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg
     1861 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
     1921 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     1981 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     2041 tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag
     2101 ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca
     2161 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     2221 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc
     2281 ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct
     2341 gccctgactt ttatgcccag cccgagctca ccgctggctg tggccttatg ggcccagggt
     2401 cctgaatgcc ctccttcact cttcttcttc cccacccgca tgccccatcc tcggacccaa
     2461 aactgtagca gtgaattgat gagtccagcg caaagcacgc cccacacaca cagggctcac
     2521 tgcggagcac gtcaaccacc actgtctccg tggcaaccgg gggtttaatt aacaggctgc
     2581 agccccatca ccatagcaac agcaccccaa cagcccctca ttcctctgca gactcagtcc
     2641 atcttccctc tttcccaatc cggaccacgc gtgcggccgc aggaacccct agtgatggag
     2701 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2761 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc
     2821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4501 ttttcaatat tattgaagca ttt
//
