LOCUS       sequence-473517-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0128h 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 5126)
  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 5126)
  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: AiP12352 -
            pAAV-AiE0128h-minRho-SYFP2-10aa-H2B...
FEATURES             Location/Qualifiers
     source          1..5126
                     /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      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(736..738,739..741,742..1452)
                     /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(781..802)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1042..1061)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1389..1410
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1483..1860
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1953..1973)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2160..2271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2389..2529
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2389..2518
                     /label=AAV2 ITR
     rep_origin      2604..3059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2691..2710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2901..2922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3076..3095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3195..3217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3255..3273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3341..3445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3446..3514,3515..4306)
                     /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(3664..3683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4966..4985
                     /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 gcgtggtcta gtatcatgag gaggatctca aagcagggaa
      181 gggaagccag ctgagggacc cctgagagga gatcagggga taaatacttt gcaacagaat
      241 ccccatggga aacatgacaa aattagtaac aggaatggag cctccctcag aggatacatg
      301 aatcgttgag ttgaaaataa ccacaaggtc atcaaatcca gctgccacca actgcctgaa
      361 tattttcaac aaatttttga acaagcagtc ctccagcctc ctccagctgc atccagcagc
      421 tcagtgctcc acctgacagc ctcttctgat agcgtgaagg ctcagaaagc ccctgtgggg
      481 cccagcacat cctggggcag gcctctagtg ggcagagggc agcatactgg ggtctggatc
      541 taacacactc ctccatccag gtggtatatt tataggagct cgattcagcc gggagcttag
      601 ggaggggagg tcacttcata agggcttggg gggggagttg gagccacgag tcgtccagcc
      661 ggagccccgt gtggctgtgc tccggcctca gaagcatccc cggatccttc gaagctagcg
      721 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      781 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      841 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
      901 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
      961 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1021 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1081 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1141 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1201 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1261 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1321 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     1381 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1441 gagctgtaca agtccggact cagatctgga ggctccggag gcccagagcc agcgaagtct
     1501 gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc
     1561 aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtataa ggttctgaag
     1621 caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa ctcgtttgtg
     1681 aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta caacaagcgc
     1741 tcgaccatca cctccaggga gatccagacg gccgtgcgcc tgctgctgcc tggggagttg
     1801 gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag cgctaagtaa
     1861 tgagtcgacg gcgcgcccct gcagggaatt cgatatcata atcaacctct ggattacaaa
     1921 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1981 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2041 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2101 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2161 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2221 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2281 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2341 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2401 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2461 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2521 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2581 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2941 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3121 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3181 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3241 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4861 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4921 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4981 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5041 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5101 cttttgctgg ccttttgctc acatgt
//
