LOCUS       sequence-473515-        5396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0082h 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 5396)
  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 5396)
  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: AiP12351 -
            pAAV-AiE0082h-minRho-SYFP2-10aa-H2B...
FEATURES             Location/Qualifiers
     source          1..5396
                     /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      1000..1009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1006..1008,1009..1011,1012..1722)
                     /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(1051..1072)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1312..1331)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1659..1680
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1753..2130
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2223..2243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2430..2541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2659..2799
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2659..2788
                     /label=AAV2 ITR
     rep_origin      2874..3329
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2961..2980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3171..3192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3346..3365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3465..3487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3525..3543)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3611..3715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3716..3784,3785..4576)
                     /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(3934..3953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4747..5335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5236..5255
                     /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 gcgtggtcct ttggccggca ggagcagcct gattttagtg
      181 actcgttcag aagggaggtg cagctggaag gttgtgttta ttaaagccca agtgactgga
      241 ggtaaatgca ggtgtgttca ggaaatgtag ctggttcacc tctcagtagg cacaaggcca
      301 aagtcagccg acttgcagat tgtaatgccg gttctcccac ccagaggtta tttgagttca
      361 tccagcactg cagctgagca gaatgctgca gtcagggtga cacaatgcaa actggagcca
      421 tgtcctcaag ggcatgccaa tcaaaccaac tttaaccttc gctggtttgt ggctgctgac
      481 ctagagagga atttgttata tggctgaggt cctgtcctac ctaagccaga ctccagtgca
      541 aaaaaaaatc ctgtcagatg cgcctcttct tctctgtctc catctccctc tctgcctccc
      601 gtttcccctt tactctttac tgttagttgg cctggggttc ctgggctgaa gtggagtttc
      661 ctcctgtctc cgtgtgtttg ctgctctggt ttgctcttct tccagggccc cttggtaaaa
      721 gaagagctgc tgacagggta gagtggaggt ggggaagtga ccaccagact gggagtgatg
      781 taggttgtgg tcatcaatga tgtaggttgc tgtcaccagg acactcaaat cagctgtgac
      841 cagaggagct cgattcagcc gggagcttag ggaggggagg tcacttcata agggcttggg
      901 gggggagttg gagccacgag tcgtccagcc ggagccccgt gtggctgtgc tccggcctca
      961 gaagcatccc cggatccttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
     1021 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1081 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1141 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1201 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1261 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1321 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1381 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1441 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1501 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1561 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1621 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1681 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatctgga
     1741 ggctccggag gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     1801 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     1861 tattccatct atgtgtataa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     1921 aaggccatgg gcatcatgaa ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag
     1981 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg
     2041 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     2101 gccatcacca agtacaccag cgctaagtaa tgagtcgacg gcgcgcccct gcagggaatt
     2161 cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     2341 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2401 gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt
     2461 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2521 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2581 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg
     2641 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2701 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2761 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2821 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2941 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3061 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3181 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3241 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3301 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3361 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3421 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3481 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3541 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3601 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3661 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3721 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3781 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3841 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3901 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3961 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4021 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4081 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4141 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4201 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4261 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4321 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4381 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4441 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4501 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4561 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4621 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4741 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4861 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
