LOCUS       sequence-446922-        5559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0961h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  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 5559)
  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: AiP13292 -
            pAAV-AiE0961h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5559
                     /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      1025..1034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1031..1033,1034..1036,1037..1747)
                     /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(1337..1356)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1684..1705
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1757..1813
                     /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             1826..1891
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1922..2299
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2386..2406)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2593..2704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2822..2962
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2822..2951
                     /label=AAV2 ITR
     rep_origin      3037..3492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3334..3355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3509..3528)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3628..3650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3688..3706)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3774..3878
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3879..3947,3948..4739)
                     /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(4097..4116)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4910..5498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5399..5418
                     /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 gcgtacacac acattcctac acacacatat tcactcacac
      181 actcatacac tcacatacac atgtatctca cacaacccac actctcacac acattcacac
      241 actcaataac tcacacactg acaggaatat tttctcattt cacccacaca aaaaaaccac
      301 tagttttgaa acaattagat tccctgagaa ttgatgtctc cctgacacaa tcatacttta
      361 aaaggcaggt cttaagtgaa gggatgactt tatcgtttaa ttttatcaat agttgggcgt
      421 aagtaaaact ttcttgaata ggaacttaca gtactgatga gaaagaccca caggactgtt
      481 catgaataaa catgaatgtg cctattgttt cgtgctgcac acaatcctat tatagaaaaa
      541 tattcaaatt atatagatac tctattataa aagaaaacct tatgtgacat gggcttattt
      601 cagggctctg tcaaaacagg gctgtcactt taatgtcaaa aaagggagtg aattttctgc
      661 atctacctac cccactcttc ccggcaatgg ctcctgtcat cttcagggac ctgaactgac
      721 ttttctggag gtggctctga agagactaga acaagcatca cctctggagg gaaacttgca
      781 acatcttttt ccttcccttt ctctcttctc tacttttctg tgttcttcct cctggaaatt
      841 ccctaacatt tcctgctctg tttagtcagt tggccatcaa gctgtctgca agggttttcc
      901 tttctagcaa tgactttgct gctgagaccc tgagctcggg ctgggcataa aagtcagggc
      961 agagccatct attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc
     1021 ggtcgccacc atggtctcca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1081 cgagctggac ggcgatgtca acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1141 tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc
     1201 ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga
     1261 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1321 caccatcttc ttcaaagacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1381 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1441 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
     1501 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     1561 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1621 cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1681 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1741 gtacaagggc tctggtgcta ccaacttctc actgttgaaa caggcagggg atgtagagga
     1801 gaatccaggg cctggtgcta gtggagacta caaagaccat gacggagatt ataaagatca
     1861 tgacatcgat tacaaggatg acgatgacaa gtccggactc agatctggag gctccggagg
     1921 cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa
     1981 ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc aaggagagct attccatcta
     2041 tgtgtacaag gttctgaagc aggtccaccc tgacaccggc atttcgtcca aggccatggg
     2101 catcatgaat tcgtttgtga acgacatttt cgagcgcatc gcaggagagg cttcccgcct
     2161 ggcgcattac aacaagcgct cgaccatcac ctcccgggag atccagacgg ccgtgcgcct
     2221 gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa
     2281 gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga attcgatatc ataatcaacc
     2341 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2401 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2461 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     2521 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg
     2581 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2641 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2701 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2761 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     2821 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2881 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2941 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3001 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3241 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3361 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     3421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3481 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3541 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3601 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3661 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3721 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3901 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3961 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4021 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4081 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4141 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4201 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4261 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4321 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4381 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4441 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4501 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4561 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4621 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4681 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4741 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5521 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
