LOCUS       sequence-446825-        5128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1026h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal MSNs (DMS>DLS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5128)
  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 5128)
  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: AiP13868 -
            pAAV-AiE1026h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5128
                     /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      594..603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(600..602,603..605,606..1316)
                     /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(906..925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1253..1274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1326..1382
                     /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             1395..1460
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1491..1868
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1955..1975)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2162..2273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2391..2531
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2391..2520
                     /label=AAV2 ITR
     rep_origin      2606..3061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2693..2712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2903..2924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3078..3097)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3197..3219
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3257..3275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3343..3447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3448..3516,3517..4308)
                     /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(3666..3685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4479..5067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4968..4987
                     /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 gcgttctttt ccatgttaga agcaaagttt taaaaaatac
      181 tctttatgct tttatgcaaa tgttatgtga tttttaaaag tattatgcat aaagacaata
      241 aaaaattaaa agtaaatact tcttttttca ttagcaggtg ttcagtaatg tgtcaaatct
      301 tgatatacct aatttgtctt ctttcaactt tacttttgga tttcctgatt ccaatttttc
      361 aaaagttgac agacatcatg ttattgtcac taattcctcc cactcagctc tcagaggttc
      421 attgttctga gggagaaatt gtcatctccc cttttccctc cactggctca gtttaagaaa
      481 gtgagtcaga ttacgggtct gagctcgggc tgggcataaa agtcagggca gagccatcta
      541 ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca
      601 tggtctccaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      661 gcgatgtcaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      721 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      781 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
      841 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      901 tcaaagacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      961 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1021 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     1081 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     1141 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1201 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1261 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggct
     1321 ctggtgctac caacttctca ctgttgaaac aggcagggga tgtagaggag aatccagggc
     1381 ctggtgctag tggagactac aaagaccatg acggagatta taaagatcat gacatcgatt
     1441 acaaggatga cgatgacaag tccggactca gatctggagg ctccggaggc ccagagccag
     1501 cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga
     1561 aaggcggcaa gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtataagg
     1621 ttctgaagca ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaact
     1681 cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca
     1741 acaagcgctc gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg
     1801 gggagttggc caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg
     1861 ctaagtaatg aggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     1921 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1981 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2041 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     2101 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     2161 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2221 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2281 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2341 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     2401 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2461 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2521 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2581 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2641 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     2701 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2761 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2821 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2881 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     2941 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3001 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3061 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3121 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3181 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3241 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3301 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3361 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3421 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3481 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3541 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3601 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3661 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3721 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3781 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3841 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3901 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3961 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4021 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4081 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4141 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4321 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5101 gccttttgct ggccttttgc tcacatgt
//
