LOCUS       sequence-472510-        4804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0447h is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4804)
  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 4804)
  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: AiP14451:
            pAAV-AiE0447h-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..4804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      412..421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(418..420,421..423,424..1134)
                     /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(463..484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(724..743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1071..1092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1227..1247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1446..1557
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1675..1815
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1675..1804
                     /label=AAV2 ITR
     rep_origin      1890..2345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1977..1996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2187..2208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2362..2381)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2481..2503
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2541..2559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2627..2731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2732..2800,2801..3592)
                     /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(2950..2969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3763..4351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4252..4271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4413..4542
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tcttgctagt tcctcctaca ttcctattgt tcgtgttaaa gatgactttt ctattcacct
       61 gtgtaagtct tgctgcctcc tacactgatt aatttagcta gattgaaaag gaggactagt
      121 gcttcctcca taaaactcca attaatcaat cattttagaa cttgagctac tttgtgtggt
      181 ggtatttgtg gaaagagcag gaatattgct gtcagtcctg agaggtggca tttactggca
      241 gatagacctc agacaagtga ttttactcct gtaagcctga atttgtttat agtaccccaa
      301 ctcataagat tggtgagaga gctcgggctg ggcataaaag tcagggcaga gccatctatt
      361 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
      421 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      481 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      541 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      601 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      661 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      721 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      841 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
      901 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1021 ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagtc
     1141 gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     1201 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1261 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1321 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     1381 acaggggctc ggctgttggg cactgacaat tccgtggctc gaggctcagg acgagagatc
     1441 ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1501 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1561 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1621 ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac
     1681 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     1741 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     1801 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1861 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     1921 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     1981 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2041 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2101 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2161 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2221 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     2281 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     2341 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2401 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2461 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2521 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3121 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3241 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3901 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4141 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4381 cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct
     4441 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     4501 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgttatg
     4561 ctgtggcctg gggatgggag ggagaatcca caggctgtgg ggatatttca taggcttcat
     4621 ctaggcatta aatgaggaaa tgaatgcaaa agtgcccagc acagtgcctg acacataaaa
     4681 gcaccaataa gcttcagctg tcttccccct tcttcaaact acttaatttt ctgtgaaact
     4741 aacacttctt ttctaattgc ccttgacttc acattcttta gcccattctt cgttactaat
     4801 ccat
//
