LOCUS       sequence-470664-        5095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0975h 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 5095)
  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 5095)
  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: AiP13459 -
            pAAV-AiE0975h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(112..903,904..972))
                     /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     735..754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(973..1077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1145..1163
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1201..1223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1323..1342
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1359..1814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1496..1517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1708..1727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1889..2029
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1900..2029
                     /label=AAV2 ITR
     polyA_signal    2147..2258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2445..2465
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2552..2929)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(2960..3025)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3038..3094)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(3104..3814,3815..3817,3818..3820))
                     /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(3146..3167)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3495..3514
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3817..3826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4257..4386
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
       61 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1081 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     1141 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1201 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1261 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1321 tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt
     1381 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat
     1441 cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt
     1501 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1561 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     1621 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1681 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc
     1741 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1801 gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc
     1861 gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact
     1921 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     1981 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcg
     2041 gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     2101 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     2161 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     2221 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct cgagccacgg
     2281 aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga
     2341 gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg aaagccatac
     2401 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     2461 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat
     2521 gatatcgaat tcgcggccgc ggcgcgcctc attacttagc gctggtgtac ttggtgatgg
     2581 ccttagtacc ctcggacacg gcgtgcttgg ccaactcccc aggcagcagc aggcgcacgg
     2641 ccgtctggat ctcccgggag gtgatggtcg agcgcttgtt gtaatgcgcc aggcgggaag
     2701 cctctcctgc gatgcgctcg aaaatgtcgt tcacaaacga attcatgatg cccatggcct
     2761 tggacgaaat gccggtgtca gggtggacct gcttcagaac cttgtacaca tagatggaat
     2821 agctctcctt gcggctgcgc ttgcgcttct tgccgccttt cttctgcgcc ttagtcaccg
     2881 ccttcttgga gccctttttc ggggcgggag cagacttcgc tggctctggg cctccggagc
     2941 ctccagatct gagtccggac ttgtcatcgt catccttgta atcgatgtca tgatctttat
     3001 aatctccgtc atggtctttg tagtctccac tagcaccagg ccctggattc tcctctacat
     3061 cccctgcctg tttcaacagt gagaagttgg tagcaccaga gcccttgtac agctcgtcca
     3121 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     3181 cgttggggtc tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca
     3241 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     3301 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3361 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3421 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3481 tcacctcggc gcgggtcttg tagttgccgt cgtctttgaa gaagatggtg cgctcctgga
     3541 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3601 gggcgaagca ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     3661 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3721 cctcgccgga cacgctgaac ttgtggccgt tgacatcgcc gtccagctcg accaggatgg
     3781 gcaccacccc ggtgaacagc tcctcgccct tggagaccat ggtggcgacc ggtagcgcta
     3841 gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt
     3901 ttatgcccag cccgagctca agaaaggcct ccaatctccc actcaaactt taaccacgta
     3961 ttttcaggag atcctcagag agtctcacag ccatcccaaa agtgccacgg catgttcttc
     4021 cactccaggc tctgtccctt gccaagtcac tttcaaagct gggccccacc actgtctctg
     4081 tagctacgac aggctgagag cgacagtaac cctctgttcc caaagctata agccactgag
     4141 gtgccaacat gaggctgtgc ttcctgcaga tgccactgag ttagttacag gcacagaact
     4201 ggggaaaatc acttcttcca ttcagcgctt ttctcactgg gaacgcgtgc ggccgcagga
     4261 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4321 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4381 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgtta
//
