LOCUS       sequence-453374-        4782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0779m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal direct pathway MSNs 
            (D1-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4782)
  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 4782)
  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: AiP15747:
            pAAV-AiE0779m_3xC2-minBG-mScarlet-WP...
FEATURES             Location/Qualifiers
     source          1..4782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      821..830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             827..1522
                     /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(960..978)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1228..1247
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1615..1635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1822..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2045..2185
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2045..2174
                     /label=AAV2 ITR
     rep_origin      2260..2715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2347..2366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2557..2578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2732..2751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2851..2873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2911..2929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2997..3101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3102..3170,3171..3962)
                     /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(3320..3339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4133..4721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4622..4641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagtt tgagacaaat taaggaaaag
      181 cgggttggga ggagtctgag gctgtgggtg gctgcctcga gaggctgctc acaacctgac
      241 atgtttcaga tcttttgaca cggtcatgca agcaacagaa acttatatgg tttttagtct
      301 cccaggtagc agggaaatga tgatgtggga ggattagggg ggggaaaata agtttgagac
      361 aaattaagga aaagcgggtt gggaggagtc tgaggctgtg ggtggctgcc tcgagaggct
      421 gctcacaacc tgacatgttt cagatctttt gacacggtca tgcaagcaac agaaacttat
      481 atggttttta gtctcccagg tagcagggaa atgatgatgt gggaggatta ggggggggaa
      541 aataagtttg agacaaatta aggaaaagcg ggttgggagg agtctgaggc tgtgggtggc
      601 tgcctcgaga ggctgctcac aacctgacat gtttcagatc ttttgacacg gtcatgcaag
      661 caacagaaac ttatatggtt tttagtctcc caggtagcag ggaaatgatg atgtgggagg
      721 attagggggg ggaaaatgag ctcgggctgg gcataaaagt cagggcagag ccatctattg
      781 cttacatttg cttctgggat ccagatcttt cgaagctagc gccaccatgg tgagcaaggg
      841 cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct ccatgaacgg
      901 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      961 caagctgaag gtgaccaagg gtggccccct gcccttctcc tgggacatcc tgtcccctca
     1021 gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc gacatccccg actactataa
     1081 gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc
     1141 cgtgaccgtg acccaggaca cctccctgga ggacggcacc ctgatctaca aggtgaagct
     1201 ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa tgggctggga
     1261 agcgtccacc gagcggttgt accccgagga cggcgtgctg aagggcgaca ttaagatggc
     1321 cctgcgcctg aaggacggcg gccgctacct ggcggacttc aagaccacct acaaggccaa
     1381 gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca tcacctccca
     1441 caacgaggac tacaccgtgg tggaacagta cgaacgctcc gagggccgcc actccaccgg
     1501 cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca
     1561 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1621 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1681 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1741 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1801 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgcacgtgcg gaccgagcgg
     2041 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2101 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2161 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2221 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2281 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2341 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2401 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2461 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2521 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2581 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2641 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2701 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2761 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2821 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2881 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2941 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3001 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3661 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3721 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3781 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3841 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3901 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3961 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4021 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4261 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
