LOCUS       sequence-459395-        4781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0452h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal indirect pathway MSNs (D2-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4781)
  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 4781)
  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: AiP15745: pAAV-AiE0452h-minBG-mScarlet-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4781
                     /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      814..823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             820..1515
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(953..971)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1221..1240
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1608..1628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1815..1926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2044..2184
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2044..2173
                     /label=AAV2 ITR
     rep_origin      2259..2714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2556..2577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2731..2750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2850..2872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2910..2928)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2996..3100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3101..3169,3170..3961)
                     /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(3319..3338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /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 gcgtagggag gaactaaagt ccattcatat gtccatattc
      181 tcaatttggg aaaaaatgtg gctttgtctt catcctcttt tcctgagtgt ataaggccag
      241 aagccaaagg caagagcagg acattccagg agcctttatg attgctctaa gtcacatctg
      301 ctctcgagct cccattccac tgtgaaaaat acctagtctt gaactttgga gttccaaatt
      361 ccacatgatc gacttcctca aggcttttca cttgacgaac aaattgcatg ccaagaaatc
      421 gtcaaaattc tatcaagctt aattatgcaa tttctgtcta taaatgacat acctcctgta
      481 ccctatgaca cccacctatg cactaataag atgtgacagc tgcatcacca acaaagcctc
      541 ccccgcctgc cattctcttc ggaggtgagc acctcctgct cgggaggaac tgacagcccc
      601 tttcagaaat gtgaaggcag tgaagtccca aaccagaaac agtgtgacca cctaggctca
      661 cccaaggacc gtcctgtgct tctgggcaac caatggatgt agggggttag gaggcagtgg
      721 ggattcgtca gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctgg gatccagatc tttcgaagct agcgccacca tggtgagcaa gggcgaggca
      841 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag
      901 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      961 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg
     1021 tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc
     1081 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc
     1141 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc
     1201 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc
     1261 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc
     1321 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc
     1381 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag
     1441 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg
     1501 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     1561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1681 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1741 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1801 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1861 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1921 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1981 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     2041 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2101 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2161 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2221 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2401 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2461 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2581 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2701 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2761 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2821 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2881 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2941 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3001 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3301 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3361 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3421 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3481 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3541 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3601 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3661 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3721 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3781 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3841 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3901 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3961 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4741 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
