LOCUS       sequence-469440-        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"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1464..1473
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1470..2165
                     /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(1603..1621)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1871..1890
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2258..2278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2465..2576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2694..2834
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2694..2823
                     /label=AAV2 ITR
     rep_origin      2909..3364
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2996..3015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3206..3227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3381..3400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3500..3522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3560..3578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3646..3750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3751..3819,3820..4611)
                     /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(3969..3988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtagggag gaactaaagt ccattcatat gtccatattc tcaatttggg
      841 aaaaaatgtg gctttgtctt catcctcttt tcctgagtgt ataaggccag aagccaaagg
      901 caagagcagg acattccagg agcctttatg attgctctaa gtcacatctg ctctcgagct
      961 cccattccac tgtgaaaaat acctagtctt gaactttgga gttccaaatt ccacatgatc
     1021 gacttcctca aggcttttca cttgacgaac aaattgcatg ccaagaaatc gtcaaaattc
     1081 tatcaagctt aattatgcaa tttctgtcta taaatgacat acctcctgta ccctatgaca
     1141 cccacctatg cactaataag atgtgacagc tgcatcacca acaaagcctc ccccgcctgc
     1201 cattctcttc ggaggtgagc acctcctgct cgggaggaac tgacagcccc tttcagaaat
     1261 gtgaaggcag tgaagtccca aaccagaaac agtgtgacca cctaggctca cccaaggacc
     1321 gtcctgtgct tctgggcaac caatggatgt agggggttag gaggcagtgg ggattcgtca
     1381 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
     1441 gatccagatc tttcgaagct agcgccacca tggtgagcaa gggcgaggca gtgatcaagg
     1501 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     1561 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1621 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1681 gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
     1741 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     1801 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     1861 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     1921 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     1981 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     2041 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     2101 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     2161 acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     2221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2341 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     2401 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     2461 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2521 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2581 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2641 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2701 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2761 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2821 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2881 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2941 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     3001 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3061 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3121 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3181 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3241 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3301 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3361 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3541 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3601 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3661 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3721 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3781 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3841 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3901 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3961 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4021 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4081 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4141 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4201 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4261 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4321 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4381 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4441 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4501 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4561 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4621 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4681 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4741 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
