LOCUS       sequence-459525-        5104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0873m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal cholinergic 
            interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5104)
  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 5104)
  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: AiP15385:
            pAAV-AiE0873m_3xC2-minBG-mScarlet-WP...
FEATURES             Location/Qualifiers
     source          1..5104
                     /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      1137..1146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1143..1838
                     /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(1276..1294)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1544..1563
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1931..1951)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2138..2249
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2367..2507
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2367..2496
                     /label=AAV2 ITR
     rep_origin      2582..3037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2669..2688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2879..2900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3054..3073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3173..3195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3233..3251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3319..3423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3424..3492,3493..4284)
                     /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(3642..3661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4455..5043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4944..4963
                     /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 gcgtcatgga tttctccaaa gctttgtagg gcagaactct
      181 gaatgtttat ttctctgatg aatccttaat gaaagtggcc agccatcctg gggagaacaa
      241 gccttagttc ccactgagaa ggatagcaag cagcataaga gtagctgatg tcaaaggcta
      301 ttaatgcaaa tgaaaattat aattcctcat caagatctgc tcccagacag gctaatcatt
      361 gtgccacctg tttacagaaa cacactgcta atcagtcagt aaaatctttt aagtttctgt
      421 gcttctctgt gtacgatttt tagcctccat ggatttctcc aaagctttgt agggcagaac
      481 tctgaatgtt tatttctctg atgaatcctt aatgaaagtg gccagccatc ctggggagaa
      541 caagccttag ttcccactga gaaggatagc aagcagcata agagtagctg atgtcaaagg
      601 ctattaatgc aaatgaaaat tataattcct catcaagatc tgctcccaga caggctaatc
      661 attgtgccac ctgtttacag aaacacactg ctaatcagtc agtaaaatct tttaagtttc
      721 tgtgcttctc tgtgtacgat ttttagcctc catggatttc tccaaagctt tgtagggcag
      781 aactctgaat gtttatttct ctgatgaatc cttaatgaaa gtggccagcc atcctgggga
      841 gaacaagcct tagttcccac tgagaaggat agcaagcagc ataagagtag ctgatgtcaa
      901 aggctattaa tgcaaatgaa aattataatt cctcatcaag atctgctccc agacaggcta
      961 atcattgtgc cacctgttta cagaaacaca ctgctaatca gtcagtaaaa tcttttaagt
     1021 ttctgtgctt ctctgtgtac gatttttagc ctcgagctcg ggctgggcat aaaagtcagg
     1081 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcca
     1141 ccatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     1201 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1261 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     1321 acatcctgtc ccctcagttc atgtacggct ccagggcctt caccaagcac cccgccgaca
     1381 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1441 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     1501 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     1561 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     1621 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     1681 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     1741 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     1801 gccgccactc caccggcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
     1861 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1921 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1981 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     2041 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2101 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     2161 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2221 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2281 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     2341 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     2401 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2461 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     2521 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     2581 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2641 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2701 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2761 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2821 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2881 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     2941 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     3001 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3061 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3121 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3181 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3241 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3301 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3361 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3421 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3481 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3541 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3601 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3661 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3721 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3781 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3841 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3901 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3961 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4021 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4081 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4141 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4201 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4261 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4321 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4381 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4441 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4501 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4561 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4621 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4681 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4741 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4801 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4861 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4921 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4981 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5041 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5101 atgt
//
