LOCUS       sequence-446934-        4776 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 4776)
  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 4776)
  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: AiP15827 - pAAV-AiE0780m-minCMV-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             8..16
                     /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   250..379
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(441..1029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(521..540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1200..1991,1992..2060))
                     /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     1823..1842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2061..2165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2233..2251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2289..2311)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2411..2430
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2447..2902)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2584..2605)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2796..2815
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2977..3117
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2988..3117
                     /label=AAV2 ITR
     polyA_signal    3235..3346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3533..3553
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3646..4356,4357..4359,4360..4362))
                     /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(3688..3709)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4037..4056
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4296..4317
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4359..4368
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4430..4468)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 atcagacacc tccttagctg ggagggtggg agagtgtcgg aggaatgagt cagttgtcag
       61 cttcgctgcg ggctcatctc tgagcctcag gtacctgggc ccggctccag actttatctg
      121 tgtctccttg ttctatggct gaacgaagga gtgctggagg ggtctcgagg tcataccagc
      181 cttagatgcc atgttggcat ggctgtgctc cagtgaacct gtagtttgtg aaacaacgcg
      241 tgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
      301 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga
      361 gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2161 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2221 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2281 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2341 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2401 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt
     2461 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2521 accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt
     2581 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2641 aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg
     2701 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2761 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2821 gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2881 aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc
     2941 acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg cgcgctcgct
     3001 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     3061 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     3121 gccgctcggt ccgcacgtga gatctcatgc ctgctattgt cttcccaatc ctcccccttg
     3181 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     3241 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     3301 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga agatctctcg
     3361 agccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
     3421 ggcgatgagt tccgccgtgg caagaactaa ccaggattta tacaaggagg agaaaatgaa
     3481 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     3541 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     3601 ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg acttacttgt acagctcgtc
     3661 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     3721 ctcgttgggg tctttgctca gcttggactg gtagctcagg tagtggttgt cgggcagcag
     3781 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc
     3841 gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc
     3901 ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     3961 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     4021 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     4081 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     4141 gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg agggtgggcc agggcacggg
     4201 cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc
     4261 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     4321 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc
     4381 tagcttcgaa agatctggat ccccaggcga tctgacggtt cactaaacca gctctgctta
     4441 tataggcctc ccaccgtaca cgcctacctc tgagctcagc ctctcgggag tatctttttc
     4501 tccatctttg agtatctgct ttctatcctt tctgttcatt tgttgttgtt gctgtgattt
     4561 ctgttgccgt tgctgcaggg ttttttcgtg gctctccacc tgcacatttg taactgacaa
     4621 gagggtgtga agaagaatga gtgatgggaa gcgctaaaag ttaatgcact cagcaaacag
     4681 agaaatccta accagtaata agcataattg ctcttcaaaa tccacccaaa gccctcgccc
     4741 ccaacagcag gcaacaaaga ggcagaacat gttcac
//
