LOCUS       sequence-446897-        4504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0612h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal D2 MSN>D1 MSN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4504)
  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 4504)
  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: AiP12442 - pAAV-AiE0612h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4504
                     /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      516..525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(522..524,525..527,528..1238)
                     /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(567..588)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(828..847)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1175..1196
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1331..1351)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1538..1649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1767..1907
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1767..1896
                     /label=AAV2 ITR
     rep_origin      1982..2437
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2088)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2279..2300
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2454..2473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2573..2595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2633..2651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2719..2823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2824..2892,2893..3684)
                     /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(3042..3061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3855..4443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4344..4363
                     /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 gcgtttccca gccttacctt gttgcaggcc tccagggtct
      181 gcagcggggt gaggcagggt ctgggagatg cagtgaatgc aggaaggggt gtctgcctga
      241 cagccccatc cctgcagatt ttgatggctc cgtaaaaatg tctcctcata aactgtcaaa
      301 gtgtcacaaa ttaattaact gttagctaga gtgactcagt gtgggggggg tgggggagaa
      361 tggctcagtg atgctgagcg gggtggtggc ggtgctgttc tctgcgtgtg ctgccttttg
      421 atgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      481 gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg
      541 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      601 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
      661 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
      721 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      781 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      841 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      901 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      961 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1021 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1081 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
     1141 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1201 ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
     1261 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1321 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1381 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     1441 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     1501 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     1561 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1621 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1681 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     1741 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     1801 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     1861 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     1921 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     1981 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2041 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2101 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2161 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2221 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2281 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     2341 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     2401 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     2461 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2521 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2581 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2641 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3541 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4501 atgt
//
