LOCUS       sequence-459399-        4729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0780m 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 4729)
  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 4729)
  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: AiP15746: pAAV-AiE0780m-minBG-mScarlet-WPRE3-B...
FEATURES             Location/Qualifiers
     source          1..4729
                     /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             364..372
                     /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      762..771
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             768..1463
                     /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(901..919)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1169..1188
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1556..1576)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1763..1874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1992..2132
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1992..2121
                     /label=AAV2 ITR
     rep_origin      2207..2662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2294..2313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2504..2525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2679..2698)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2798..2820
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2858..2876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2944..3048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3049..3117,3118..3909)
                     /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(3267..3286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /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 gcgttgtttc acaaactaca ggttcactgg agcacagcca
      181 tgccaacatg gcatctaagg ctggtatgac ctcgagaccc ctccagcact ccttcgttca
      241 gccatagaac aaggagacac agataaagtc tggagccggg cccaggtacc tgaggctcag
      301 agatgagccc gcagcgaagc tgacaactga ctcattcctc cgacactctc ccaccctccc
      361 agctaaggag gtgtctgatg tgaacatgtt ctgcctcttt gttgcctgct gttgggggcg
      421 agggctttgg gtggattttg aagagcaatt atgcttatta ctggttagga tttctctgtt
      481 tgctgagtgc attaactttt agcgcttccc atcactcatt cttcttcaca ccctcttgtc
      541 agttacaaat gtgcaggtgg agagccacga aaaaaccctg cagcaacggc aacagaaatc
      601 acagcaacaa caacaaatga acagaaagga tagaaagcag atactcaaag atggagaaaa
      661 agatactccc gagaggctga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      721 gcttacattt gcttctggga tccagatctt tcgaagctag cgccaccatg gtgagcaagg
      781 gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg
      841 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
      901 ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc
      961 agttcatgta cggctccagg gccttcacca agcaccccgc cgacatcccc gactactata
     1021 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1081 ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc
     1141 tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg
     1201 aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg
     1261 ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc tacaaggcca
     1321 agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac atcacctccc
     1381 acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc cactccaccg
     1441 gcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     1501 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1561 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     1621 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     1681 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     1741 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1801 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1861 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1921 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     1981 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2041 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2101 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2161 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2221 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2281 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2341 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2401 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2461 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2521 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     2581 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2641 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     2701 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2761 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2821 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2881 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3061 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3121 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3181 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3241 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3301 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3361 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3421 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3481 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3541 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3601 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3661 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3721 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3781 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3841 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3901 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3961 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
