LOCUS       sequence-446940-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0873m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal cholinergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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: AiP15820 -
            pAAV-AiE0873m_3xC2-minBG-RiboL1-jGC...
FEATURES             Location/Qualifiers
     source          1..6080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..50)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     241..262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(416..435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     535..557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(595..613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        681..785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(786..854,855..1646)
                     /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(1004..1023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1817..2405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2306..2325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2467..2596
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             3295..3312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3594..3615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3721..3742)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3982..4001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5373..5393)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5580..5691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5809..5949
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5809..5938
                     /label=AAV2 ITR
     rep_origin      join(6024..6080,1..399)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
       61 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      121 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
      181 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      241 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
      301 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
      361 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
      421 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      481 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
      541 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
      601 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2461 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
     2521 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     2581 atcactaggg gttcctgcgg ccgcacgcgt catggatttc tccaaagctt tgtagggcag
     2641 aactctgaat gtttatttct ctgatgaatc cttaatgaaa gtggccagcc atcctgggga
     2701 gaacaagcct tagttcccac tgagaaggat agcaagcagc ataagagtag ctgatgtcaa
     2761 aggctattaa tgcaaatgaa aattataatt cctcatcaag atctgctccc agacaggcta
     2821 atcattgtgc cacctgttta cagaaacaca ctgctaatca gtcagtaaaa tcttttaagt
     2881 ttctgtgctt ctctgtgtac gatttttagc ctccatggat ttctccaaag ctttgtaggg
     2941 cagaactctg aatgtttatt tctctgatga atccttaatg aaagtggcca gccatcctgg
     3001 ggagaacaag ccttagttcc cactgagaag gatagcaagc agcataagag tagctgatgt
     3061 caaaggctat taatgcaaat gaaaattata attcctcatc aagatctgct cccagacagg
     3121 ctaatcattg tgccacctgt ttacagaaac acactgctaa tcagtcagta aaatctttta
     3181 agtttctgtg cttctctgtg tacgattttt agcctcgagc tcgggctggg cataaaagtc
     3241 agggcagagc catctattgc ttacatttgc ttctgggatc cacccgccac catgcatcat
     3301 caccatcatc acacgcgtcg caagaagacc ttcaaggagg tggccaccgc cgtgaagatc
     3361 atcgccatgc tgatgggcct gaagatcaac gtctatatca aggccgacaa gcagaagaac
     3421 ggcatcaagg cgaacttcca catccgccac aacatcgagg acggcggcgt gcagctcgcc
     3481 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3541 tacctgagcg tggagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     3601 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     3661 ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     3781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3841 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     3961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4081 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     4141 gcagaataca aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
     4201 aaggagctgg ggacggtgat gcggtctctc ggacacaacc ccacagaagc agagctgcag
     4261 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     4321 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     4381 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     4441 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     4501 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagggg
     4561 ggcagcggtg ggtctggagg gactgggggt agcggtggta gcggtgggac agggggaagc
     4621 ggcggtagtg gtggaacaag cagcaaagtc tcacgcgaca ccctgtacga ggcggtgcgg
     4681 gaagtcctgc acgggaacca gcgcaagcgc cgcaagtttc tggagacggt ggagctgcag
     4741 atcagcctga agaactacga ccctcagaag gacaaacgtt tctcgggcac cgtcaggctc
     4801 aagtccaccc cacgccccaa gttctcggtg tgcgttctgg gggaccagca gcactgtgat
     4861 gaagccaagg ccgtggatat cccccacatg gacatcgagg cgctcaagaa acttaacaaa
     4921 aacaagaagt tggtcaagaa gctggctaag aagtacgatg cctttttggc ctctgagtct
     4981 ctgattaagc agatcccacg tatcctgggc ccaggcctaa acaaggctgg caagttcccc
     5041 tccctgctga cacacaatga aaacatggtg gccaaagtgg atgaggtgaa atcgacaatc
     5101 aagttccaga tgaagaaggt gctgtgtttg gccgtcgctg ttggccacgt gaagatgacc
     5161 gatgatgagc tagtctacaa cattcatctg gctgtcaatt tcttggtgtc cttgcttaag
     5221 aaaaactggc aaaacgtgcg ggctctgtac atcaagagca ccatgggcaa gccccagcgt
     5281 ctgtattagg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     5341 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5401 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5461 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     5521 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     5581 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5641 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5701 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5761 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     5821 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     5881 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5941 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6001 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6061 ggtggttacg cgcagcgtga
//
