LOCUS       sequence-459477-        5092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0949h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal neuron subset.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Vargas S, Lerma MN, Taskin N, Weed N, Laird WD, Bishaw YM, Bendrick 
            JL, Gore BB, Ben-Simon Y, Opitz-Araya X, Martinez RA, Way SW, 
            Thyagarajan B, Otto S, Sanchez REA, Alexander JR, Amaya A, Amster A,
            Arbuckle J, Ayala A, Baker PM, Barcelli T, Barta S, Bertagnolli D, 
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chong P, Clark M, Colbert K, 
            Daniel S, Dawe T, Departee M, DiValentin P, Donadio NP, Dotson NI, 
            Dwivedi D, Egdorf T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, 
            Gudsnuk K, Han W, Haradon Z, Hastings S, Helback O, Ho WV, Huang C, 
            Johnson T, Jones DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, 
            Loeffler H, Lusk NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor
            J, Mich JK, Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K,
            Oster AL, Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, 
            Potekhina L, Ransford S, Ray PL, Reding M, Rette DF, Reynoldson C, 
            Rimorin C, Sigler AR, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, 
            Sevigny JP, Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats 
            B, Taormina MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford 
            A, Dee N, Mollenkopf T, Ng L, Esposito L, Kalmbach BE, Yao S, Ariza 
            J, Collman F, Mufti S, Smith K, Waters J, Ersing I, Patrick M, Zeng 
            H, Lein ES, Kojima Y, Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic 
            B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   Neuron. 2025 May 21;113(10):1507-1524.e17. doi: 
            10.1016/j.neuron.2025.04.035.
  PUBMED    40403704
REFERENCE   2  (bases 1 to 5092)
  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"; doi: 
            "10.1016/j.neuron.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1507-1524.e17"
COMMENT     Sequence Label: AiP13640:
            pAAV-AiE0949h-minBG-SYFP2-P2A-3XFLAG...
FEATURES             Location/Qualifiers
     source          1..5092
                     /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      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(564..566,567..569,570..1280)
                     /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(870..889)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1217..1238
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1290..1346
                     /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             1359..1424
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1455..1832
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1919..1939)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2126..2237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2355..2495
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2355..2484
                     /label=AAV2 ITR
     rep_origin      2570..3025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2657..2676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2867..2888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3042..3061)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3161..3183
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3221..3239)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3307..3411
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3412..3480,3481..4272)
                     /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(3630..3649)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4443..5031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4932..4951
                     /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 gcgttcacag agaggttaaa tatcttgctg agatcacaaa
      181 ccagtaaatg atggagttga ttcaacccaa gatttgtctg attccagagc tcatgttctt
      241 aactatgcca cttaactatg ccactttttg gggtgaggca gacaaatgtg aaatgggtac
      301 taattttaca atctcacttg ttcagagtat gtgaacatgt cacatcatct gactaggctt
      361 ggtgaagtaa tgcaggtgca cactgctcat agagccagtg ggagttacat tgttattatt
      421 attagagata gtctttagca tttctatgct actttcttct tgcagagctc gggctgggca
      481 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
      541 agctagcgct accggtcgcc accatggtct ccaagggcga ggagctgttc accggggtgg
      601 tgcccatcct ggtcgagctg gacggcgatg tcaacggcca caagttcagc gtgtccggcg
      661 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
      721 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
      781 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      841 acgtccagga gcgcaccatc ttcttcaaag acgacggcaa ctacaagacc cgcgccgagg
      901 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      961 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1021 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1081 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1141 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     1201 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1261 gcatggacga gctgtacaag ggctctggtg ctaccaactt ctcactgttg aaacaggcag
     1321 gggatgtaga ggagaatcca gggcctggtg ctagtggaga ctacaaagac catgacggag
     1381 attataaaga tcatgacatc gattacaagg atgacgatga caagtccgga ctcagatctg
     1441 gaggctccgg aggcccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga
     1501 aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga
     1561 gctattccat ctatgtgtac aaggttctga agcaggtcca ccctgacacc ggcatttcgt
     1621 ccaaggccat gggcatcatg aattcgtttg tgaacgacat tttcgagcgc atcgcaggag
     1681 aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctcccgg gagatccaga
     1741 cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta
     1801 aggccatcac caagtacacc agcgctaagt aatgaggcgc gccgcggccg cgaattcgat
     1861 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1921 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1981 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     2041 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2101 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2161 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2221 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2281 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     2341 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2401 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2461 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2521 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2641 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2701 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2761 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2881 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2941 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3001 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3061 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3121 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3181 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3241 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3421 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3481 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3541 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3601 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3661 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3721 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3781 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3841 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3901 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3961 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4021 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4081 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4141 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4201 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4261 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4321 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4561 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4801 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5041 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
