LOCUS       sequence-446914-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0743m_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 5677)
  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 5677)
  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: AiP13738 -
            pAAV-AiE0743m_3xC2-minBG-tdTomato-W...
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      978..987
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             984..2414
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(2504..2524)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2711..2822
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2940..3080
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2940..3069
                     /label=AAV2 ITR
     rep_origin      3155..3610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3242..3261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3452..3473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3627..3646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3746..3768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3806..3824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3892..3996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3997..4065,4066..4857)
                     /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(4215..4234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5028..5616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5517..5536
                     /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 gcgtagcagc agctagtagt acatacaaag aaatagaaaa
      181 catgaagacc aggggagttt aattatatgg ttgcaaaaat ggttcaacat tgcaaagttg
      241 ttaatactca gtggatgggg agggagctgt gcaattatct ccacaggtac agaaaaggca
      301 cttagtaaaa tgtagccaac tctttggctt ctaaaaaagg agaaaatggt ttacaaaaac
      361 aagcctccct ggatgcttcc ttaatttgtt cttagcagca gctagtagta catacaaaga
      421 aatagaaaac atgaagacca ggggagttta attatatggt tgcaaaaatg gttcaacatt
      481 gcaaagttgt taatactcag tggatgggga gggagctgtg caattatctc cacaggtaca
      541 gaaaaggcac ttagtaaaat gtagccaact ctttggcttc taaaaaagga gaaaatggtt
      601 tacaaaaaca agcctccctg gatgcttcct taatttgttc ttagcagcag ctagtagtac
      661 atacaaagaa atagaaaaca tgaagaccag gggagtttaa ttatatggtt gcaaaaatgg
      721 ttcaacattg caaagttgtt aatactcagt ggatggggag ggagctgtgc aattatctcc
      781 acaggtacag aaaaggcact tagtaaaatg tagccaactc tttggcttct aaaaaaggag
      841 aaaatggttt acaaaaacaa gcctccctgg atgcttcctt aatttgttct tgagctcggg
      901 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      961 ctttcgaagc tagcggcgcc accatggtga gcaagggcga ggaggtcatc aaagagttca
     1021 tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     1081 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     1141 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt
     1201 acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca
     1261 agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct
     1321 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     1381 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     1441 cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc
     1501 actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     1561 actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     1621 aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc atcggcagca
     1681 ccggcagcgg cagctccggc accgcctcct ccgaggacaa caacatggcc gtcatcaaag
     1741 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     1801 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1861 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     1921 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     1981 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     2041 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     2101 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     2161 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     2221 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     2281 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     2341 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg
     2401 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     2461 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2521 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2581 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     2641 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     2701 agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2761 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2821 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2881 ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca
     2941 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3001 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3061 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3121 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     3181 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     3241 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3301 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3361 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3421 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3481 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     3541 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3601 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3661 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3721 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3781 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3841 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3901 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3961 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4021 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4081 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4141 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4201 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4261 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4321 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4381 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4441 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4501 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4561 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4621 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4681 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4741 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4801 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4861 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4921 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4981 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5041 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5101 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5161 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5221 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5281 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5341 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5401 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5461 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5521 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5581 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5641 cggttcctgg ccttttgctg gccttttgct cacatgt
//
