LOCUS       sequence-459524-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0445h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal indirect pathway MSNs 
            (D2-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  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 5555)
  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: AiP13985:
            pAAV-AiE0445h_3xC2-minBG-jRGECO1a-WP...
FEATURES             Location/Qualifiers
     source          1..5555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             952..969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             973..1005
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1009..1017,1018..1032)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1812..1831
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2382..2402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2589..2700
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2818..2958
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2818..2947
                     /label=AAV2 ITR
     rep_origin      3033..3488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3120..3139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3330..3351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3505..3524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3624..3646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3684..3702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3770..3874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3875..3943,3944..4735)
                     /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(4093..4112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4906..5494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5395..5414
                     /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 gcgtgtggag gactttgagc ttaaccctat taaatttaat
      181 ctaatatatt cagcagcctg tcacaatctt gttaaatctt tattccgtca cccaccttat
      241 tagggggccc tcccacttta gtcatcagca gattagtaag catgccattt agatgtcatc
      301 taagttaatg aacctgactg agccagagtc acatggcaga gtcacagaaa ctccttctga
      361 gctggcgtgg aggactttga gcttaaccct attaaattta atctaatata ttcagcagcc
      421 tgtcacaatc ttgttaaatc tttattccgt cacccacctt attagggggc cctcccactt
      481 tagtcatcag cagattagta agcatgccat ttagatgtca tctaagttaa tgaacctgac
      541 tgagccagag tcacatggca gagtcacaga aactccttct gagctggcgt ggaggacttt
      601 gagcttaacc ctattaaatt taatctaata tattcagcag cctgtcacaa tcttgttaaa
      661 tctttattcc gtcacccacc ttattagggg gccctcccac tttagtcatc agcagattag
      721 taagcatgcc atttagatgt catctaagtt aatgaacctg actgagccag agtcacatgg
      781 cagagtcaca gaaactcctt ctgagctggc gagctcgggc tgggcataaa agtcagggca
      841 gagccatcta ttgcttacat ttgcttctgg gatccacccg ccaccatgct gcagaacgag
      901 cttgctctta agttggctgg acttgatatt aacaagactg gaggaggttc tcatcatcat
      961 catcatcatg gtatggcaag catgactggt ggacagcaaa tgggtcggga tctgtacgac
     1021 gatgacgata aggatctcgc aacaatggtc gactcatcac gtcgtaagtg gaataaggca
     1081 ggtcacgcag tcagagctat aggtcggctg agctcccccg tggtttccga gcggatgtac
     1141 cccgaggacg gcgccctgaa gagcgagatc aagaaggggc tgaggctgaa ggacggcggc
     1201 cactacgccg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1261 gcctacatcg tggacatcaa gttggacatc gtgtcccaca acgaggacta caccatcgtg
     1321 gaacagtgcg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1381 ggaggtacag gcgggagtct ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     1441 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1501 gagggcgagg gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc
     1561 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1621 aaggcctaca ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag
     1681 ggcttcaggt gggagcgcgt gatgaacttc gaggacggcg gcattattca cgtgaaccag
     1741 gactcctccc tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg caccaacttc
     1801 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggctac tcgtgacgac
     1861 ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg
     1921 gatgggacga taacaaccaa ggagcttggg acggtgttcc gatcgctggg gcagaacccc
     1981 acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacattc
     2041 gacttccctg agttcctgac gatgatggca agaaaaatga atgacacaga cagtgaagag
     2101 gaaattcgcg aagcgttccg cgtgtttgat aaggacggca atggctacat cggcgcagca
     2161 gagcttcgcc acgtgatgac agaccttgga gagaagttaa cagatgagga ggttgatgaa
     2221 atgatcaggg tagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa
     2281 atgatgacag cgaagtgagg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     2341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2461 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     2521 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     2581 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2641 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2701 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2761 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2821 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2881 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2941 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3001 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3061 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3121 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3181 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3241 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3301 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3361 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3421 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3481 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3601 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3661 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3721 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3841 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3901 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3961 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4021 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4081 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4141 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4201 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4261 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4321 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4381 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4441 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4501 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4561 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4621 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4681 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4741 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5041 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5521 gttcctggcc ttttgctggc cttttgctca catgt
//
