LOCUS       sequence-446846-        5546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0780m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal D1 MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5546)
  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 5546)
  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: AiP15828 - pAAV-AiE0780m-HSP68-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..5546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(23..478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(160..181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     372..391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   553..693
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   564..693
                     /label=AAV2 ITR
     polyA_signal    811..922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1109..1129
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1222..1932,1933..1935,1936..1938))
                     /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(1264..1285)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1613..1632
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1872..1893
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1935..1944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     RBS             3130..3138
                     /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)"
     repeat_region   3372..3501
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3563..4151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3643..3662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4322..5113,5114..5182))
                     /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     4945..4964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5183..5287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5355..5373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5411..5433)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(5533..5546,1..6)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
       61 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
      121 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
      181 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      241 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
      301 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
      361 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
      421 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
      481 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      541 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
      601 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      661 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
      721 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
      781 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
      841 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
      901 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
      961 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
     1021 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     1081 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     1141 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
     1201 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1261 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1321 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1381 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
     1441 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
     1501 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1561 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1621 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1681 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     1741 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     1801 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1861 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1921 ctcgcccttg ctcaccatgg tggcgaccgg taggccgcgc tctgcttctg gaaggctgcg
     1981 ctccgcggcg tggatgctcc ggggaaagtt gcgctccgcg gcagggatgc tcctgggaag
     2041 gttgcgctcc gcggcaggga tgctctgggg aaggctggtc ctggccgagg atcgggaacg
     2101 cgccgctcgc tctgcttctc ttgtcttcgc ttgtctctgg atggaaccag atttggttct
     2161 gagtagctgt cagcgtctgg tgacctgctc gccgccctgc gcctttaagg agtcttcacc
     2221 ggccccgccc actctccgct gggccaatca gcgagccgga ggaggccttg gggccaggaa
     2281 tcttccagca gtttcgcgtc tggtggagct tccccgcctc ccttgagtaa tcggagttgt
     2341 gggttccgcc cttgtccaga actctccaga ggtttctggg gttcactgga gagtacggat
     2401 tcctgagggg gagggtgtgg ggaagtgctg gtgctactag tgacactgtt gctatggcga
     2461 cgcattacta aggcctgtgt ggaatggaca agaaagatca cctctagctc ggtgttgtgt
     2521 acagtttgtt gtgatttgtg gggtttcgcc aactcgcaca gttctgaata tgggggttaa
     2581 aggctaaaac ttaagggcta aaacttctcc ccgccaagtt taggagaccc agggagatgc
     2641 ctgggggcgt gtccggtgac gtgatcctct ccaatcgcgt tacaatggca gtgctgcctc
     2701 tgacctcatg gactaattta ggaactagag gctctgtccc agcacaggct caaagttgcc
     2761 gggaggggcg gggtgggggg tgggggggac cccggctgct cagtttggat gttcctggag
     2821 ctcagcctct cgggagtatc tttttctcca tctttgagta tctgctttct atcctttctg
     2881 ttcatttgtt gttgttgctg tgatttctgt tgccgttgct gcagggtttt ttcgtggctc
     2941 tccacctgca catttgtaac tgacaagagg gtgtgaagaa gaatgagtga tgggaagcgc
     3001 taaaagttaa tgcactcagc aaacagagaa atcctaacca gtaataagca taattgctct
     3061 tcaaaatcca cccaaagccc tcgcccccaa cagcaggcaa caaagaggca gaacatgttc
     3121 acatcagaca cctccttagc tgggagggtg ggagagtgtc ggaggaatga gtcagttgtc
     3181 agcttcgctg cgggctcatc tctgagcctc aggtacctgg gcccggctcc agactttatc
     3241 tgtgtctcct tgttctatgg ctgaacgaag gagtgctgga ggggtctcga ggtcatacca
     3301 gccttagatg ccatgttggc atggctgtgc tccagtgaac ctgtagtttg tgaaacaacg
     3361 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3421 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
     3481 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3541 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3601 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3661 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3721 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3781 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3841 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3901 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3961 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4021 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4081 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4141 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4201 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4261 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4321 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4381 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4441 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4501 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4561 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4621 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4681 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4741 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4801 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4861 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4921 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4981 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5041 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5101 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5161 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5221 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5281 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5341 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5401 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5461 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5521 ggcttaacta tgcggcatca gagcag
//
