LOCUS       sequence-472837-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0452h_3xC2 is an enhancer active in striatal D2-MSNs and cortical
            L5_ET neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  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 4922)
  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: AiP14460:
            pAAV-AiE0452h_3xC2-minBG-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..4922
                     /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      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(928..930,931..933,934..1644)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(973..994)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1234..1253)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1581..1602
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1737..1757)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1956..2067
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2185..2325
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2185..2314
                     /label=AAV2 ITR
     rep_origin      2400..2855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2487..2506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2697..2718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2872..2891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2991..3013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3051..3069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3137..3241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3242..3310,3311..4102)
                     /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(3460..3479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /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 gcgtgtgaaa aatacctagt cttgaacttt ggagttccaa
      181 attccacatg atcgacttcc tcaaggcttt tcacttgacg aacaaattgc atgccaagaa
      241 atcgtcaaaa ttctatcaag cttaattatg caatttctgt ctataaatga catacctcct
      301 gtaccctatg acacccacct atgcactaat aagatgtgac agctgcatca ccaacaaagc
      361 ctcccccgcc tggtgaaaaa tacctagtct tgaactttgg agttccaaat tccacatgat
      421 cgacttcctc aaggcttttc acttgacgaa caaattgcat gccaagaaat cgtcaaaatt
      481 ctatcaagct taattatgca atttctgtct ataaatgaca tacctcctgt accctatgac
      541 acccacctat gcactaataa gatgtgacag ctgcatcacc aacaaagcct cccccgcctg
      601 gtgaaaaata cctagtcttg aactttggag ttccaaattc cacatgatcg acttcctcaa
      661 ggcttttcac ttgacgaaca aattgcatgc caagaaatcg tcaaaattct atcaagctta
      721 attatgcaat ttctgtctat aaatgacata cctcctgtac cctatgacac ccacctatgc
      781 actaataaga tgtgacagct gcatcaccaa caaagcctcc cccgcctgga gctcgggctg
      841 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
      901 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      961 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1021 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1081 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1141 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1201 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1261 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1321 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1381 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1441 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1501 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     1561 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1621 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     1681 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1741 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     1801 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     1861 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     1921 tccgtggctc gaggatccac ccgagagatc ttcgactgtg ccttctagtt gccagccatc
     1981 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2041 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2101 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     2161 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2221 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2281 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     2341 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     2401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2461 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2521 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2581 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2701 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2761 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2821 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2881 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2941 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3001 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3061 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3241 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3301 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3361 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3421 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3481 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3541 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3601 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3661 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3721 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3781 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3841 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3901 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3961 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4021 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4081 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gt
//
