LOCUS       sequence-459488-        4951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0743m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal cholinergic 
            interneurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4951)
  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 4951)
  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: AiP14530: pAAV-AiE0743m_3xC2-minBG-mTFP1-WPRE3...
FEATURES             Location/Qualifiers
     source          1..4951
                     /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..1691
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     primer_bind     complement(1778..1798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1985..2096
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2214..2354
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2214..2343
                     /label=AAV2 ITR
     rep_origin      2429..2884
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2516..2535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2726..2747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2901..2920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3020..3042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3080..3098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3166..3270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3271..3339,3340..4131)
                     /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(3489..3508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4302..4890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4791..4810
                     /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 agggcgagga gaccacaatg ggcgtaatca
     1021 agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac gccttcgtga
     1081 tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac ctggaggtga
     1141 aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc gcctacggca
     1201 acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag tccttccccg
     1261 agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg aaggtgaagt
     1321 ccgacatctc catggaggag gactccttca tctacgagat acacctcaag ggcgagaact
     1381 tcccccccaa cggccccgtg atgcagaaga agaccaccgg ctgggacgcc tccaccgaga
     1441 ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg
     1501 gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag gcggtgaagc
     1561 tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac aaggactaca
     1621 acaaggtgac cgtttacgag agcgccgtgg cccgcaactc caccgacggc atggacgagc
     1681 tgtacaagta atgaggcgcg ccgcggccgc gaattcgata tcataatcaa cctctggatt
     1741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1861 cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg
     1921 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct
     1981 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2041 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2101 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2161 gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc
     2221 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2281 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     2341 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2401 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     2461 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     2521 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2581 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2641 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2701 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2761 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     2821 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     2881 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     2941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3061 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3121 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3481 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3781 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg t
//
