LOCUS       sequence-447025-        4942 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 4942)
  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 4942)
  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: AiP14202 -
            pAAV-AiE0743m_3xC2-minBG-mScarlet-W...
FEATURES             Location/Qualifiers
     source          1..4942
                     /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      975..984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             981..1676
                     /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(1114..1132)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1382..1401
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1769..1789)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1976..2087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2205..2345
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2205..2334
                     /label=AAV2 ITR
     rep_origin      2420..2875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2507..2526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2717..2738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2892..2911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3011..3033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3071..3089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3157..3261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3262..3330,3331..4122)
                     /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(3480..3499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4293..4881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4782..4801
                     /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 tagcgccacc atggtgagca agggcgaggc agtgatcaag gagttcatgc
     1021 ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     1081 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1141 ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca
     1201 ccaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt
     1261 gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc
     1321 tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg
     1381 gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg
     1441 aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct
     1501 acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct
     1561 acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac
     1621 agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     1681 tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt
     1741 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     1801 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     1861 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1921 ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg
     1981 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2041 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2101 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2161 gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     2221 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     2281 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     2341 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2401 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2461 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     2521 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2581 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     2641 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2701 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     2761 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     2821 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     2881 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2941 acacccgctg acgcgccctg tcgggcttgt ctgctcccgg catccgctta cagacaagct
     3001 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3061 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3121 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3181 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3241 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3301 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3361 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3421 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3481 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3541 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3601 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3661 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3721 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3781 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3841 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3901 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3961 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4021 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4081 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4141 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4201 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4681 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4921 tgctggcctt ttgctcacat gt
//
