LOCUS       sequence-459397-        4777 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 4777)
  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 4777)
  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..4777
                     /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      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             816..1511
                     /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(949..967)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1217..1236
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1604..1624)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1811..1922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2040..2180
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2040..2169
                     /label=AAV2 ITR
     rep_origin      2255..2710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2342..2361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2552..2573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2727..2746)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2846..2868
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2906..2924)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2992..3096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3097..3165,3166..3957)
                     /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(3315..3334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4128..4716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4617..4636
                     /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 gcgtaagttt gagacaaatt aaggaaaagc gggttgggag
      181 gagtctgagg ctgtgggtgg ctgcctcgag aggctgctca caacctgaca tgtttcagat
      241 cttttgacac ggtcatgcaa gcaacagaaa cttatatggt ttttagtctc ccaggtagca
      301 gggaaatgat gatgtgggag gattaggggg gggaaaataa gtttgagaca aattaaggaa
      361 aagcgggttg ggaggagtct gaggctgtgg gtggctgcct cgagaggctg ctcacaacct
      421 gacatgtttc agatcttttg acacggtcat gcaagcaaca gaaacttata tggtttttag
      481 tctcccaggt agcagggaaa tgatgatgtg ggaggattag gggggggaaa ataagtttga
      541 gacaaattaa ggaaaagcgg gttgggagga gtctgaggct gtgggtggct gcctcgagag
      601 gctgctcaca acctgacatg tttcagatct tttgacacgg tcatgcaagc aacagaaact
      661 tatatggttt ttagtctccc aggtagcagg gaaatgatga tgtgggagga ttaggggggg
      721 gaaaatgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      781 ttctgggatc cagatctttc gaagctagcg ccaccatggt gagcaagggc gaggcagtga
      841 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
      901 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      961 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
     1021 gctccagggc cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc
     1081 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
     1141 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
     1201 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
     1261 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
     1321 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
     1381 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
     1441 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
     1501 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     1561 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1621 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1681 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     1741 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     1801 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1981 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     2041 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2101 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2161 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2221 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     2281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     2341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2401 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2461 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2581 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     2641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2701 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2761 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2821 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2881 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2941 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3061 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3121 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3181 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3241 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3301 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3361 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3421 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3481 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3541 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3601 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3661 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3721 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3781 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3841 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3901 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3961 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4021 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4261 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4501 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4741 cggttcctgg ccttttgctg gccttttgct cacatgt
//
