LOCUS       sequence-459527-        4859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0780m_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 4859)
  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 4859)
  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: AiP15649: pAAV-AiE0780m_3xC2-minBG-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..4859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     RBS             203..211
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             414..422
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             625..633
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      871..880
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(877..879,880..882,883..1593)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(922..943)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1183..1202)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1530..1551
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1686..1706)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1893..2004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2122..2262
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2122..2251
                     /label=AAV2 ITR
     rep_origin      2337..2792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2424..2443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2634..2655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2809..2828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2928..2950
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2988..3006)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3074..3178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3179..3247,3248..4039)
                     /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(3397..3416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4210..4798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4699..4718
                     /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 gcgtagcccg cagcgaagct gacaactgac tcattcctcc
      181 gacactctcc caccctccca gctaaggagg tgtctgatgt gaacatgttc tgcctctttg
      241 ttgcctgctg ttgggggcga gggctttggg tggattttga agagcaatta tgcttattac
      301 tggttaggat ttctctgttt gctgagtgca ttaactttta gcgcttccca tcactagccc
      361 gcagcgaagc tgacaactga ctcattcctc cgacactctc ccaccctccc agctaaggag
      421 gtgtctgatg tgaacatgtt ctgcctcttt gttgcctgct gttgggggcg agggctttgg
      481 gtggattttg aagagcaatt atgcttatta ctggttagga tttctctgtt tgctgagtgc
      541 attaactttt agcgcttccc atcactagcc cgcagcgaag ctgacaactg actcattcct
      601 ccgacactct cccaccctcc cagctaagga ggtgtctgat gtgaacatgt tctgcctctt
      661 tgttgcctgc tgttgggggc gagggctttg ggtggatttt gaagagcaat tatgcttatt
      721 actggttagg atttctctgt ttgctgagtg cattaacttt tagcgcttcc catcactgag
      781 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
      841 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      901 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      961 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     1021 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     1081 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1141 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1201 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1261 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1321 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1381 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1441 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     1501 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1561 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     1621 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1681 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1741 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     1801 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1861 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     1921 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1981 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2041 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2101 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2161 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2221 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2281 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2341 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2401 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2461 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2521 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2581 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2641 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     2701 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2761 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     2821 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     2881 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2941 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3001 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3061 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3301 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3361 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3421 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3481 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3541 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3601 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3661 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3721 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3781 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3841 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3901 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3961 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4021 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4321 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4801 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
