LOCUS       sequence-470455-        4738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0130h 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 4738)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4738)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP12309 - pAAV-AiE0130h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   103..232
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(294..882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(374..393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1053..1844,1845..1913))
                     /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     1676..1695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1914..2018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2086..2104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2142..2164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2264..2283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2300..2755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2437..2458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2649..2668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2830..2970
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2841..2970
                     /label=AAV2 ITR
     polyA_signal    3088..3199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3386..3406
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3499..4209,4210..4212,4213..4215))
                     /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(3541..3562)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3890..3909
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4149..4170
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4212..4221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggtcagatct ccagtgaacc cttgaaggca acgctctgca gccctcaccc caccagcctt
       61 ctgacctcta gctgtagctg cccttcccac gcgtgcggcc gcaggaaccc ctagtgatgg
      121 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
      181 cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga
      241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1141 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2041 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2101 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2161 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2221 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2281 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     2341 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     2401 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     2461 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2521 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2581 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2641 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2701 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2761 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2821 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     2881 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2941 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tgagatctca
     3001 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     3061 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     3121 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     3181 tggcaactag aaggcacagt cgaagatctc tcgagccacg gaattgtcag tgcccaacag
     3241 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac
     3301 taaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata
     3361 caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc
     3421 agtcaatctt tcacaaattt tgtaatccag aggttgatta tgatatcgaa ttcgcggccg
     3481 cggcgcgccg tcgacttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     3541 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     3601 ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     3661 ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     3721 gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgttgta
     3781 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     3841 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     3901 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     3961 gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc
     4021 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatcagctt
     4081 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     4141 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     4201 gcccttgctc accatggtgg cgaccggtag cgctagcttc gaaagatctg gatcccagaa
     4261 gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga gctctgcttc
     4321 agccattatt tattcaaata ttttggcagc tcccattttc tgcttctttc cttgcctgtt
     4381 agttgttgat tggctgccag atgttgtgaa tttcagtgtt ctgggtatgg gattcttttt
     4441 tatttttata actgtttttg aatttcattc caggatgtgt ttaagttact tagaaatggc
     4501 ttgttctttt agagtcttgc ttctaaactt tgtttgctga aaccagagca gctgttagac
     4561 tatggctaaa attggcccat tgctaaagtt aaacccttct gattgctctg ctcaatgtcc
     4621 tatatagcgt aaggctttgc attccggatg acaggaagaa gcagcatcct cagccctgtg
     4681 tggtatctga gaattgttcc tctaatactt tggggaagct ttctcatgtg catgcact
//
