LOCUS       sequence-472512-        5084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0526h is an enhancer active in striatal MSNs and cortical Sst 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5084)
  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 5084)
  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: AiP14455:
            pAAV-AiE0526h-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..5084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(107..126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     317..338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(492..511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     611..633
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(671..689)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        757..861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(862..930,931..1722)
                     /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(1080..1099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1893..2481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2382..2401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2543..2672
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3626..3635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3632..3634,3635..3637,3638..4348)
                     /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(3677..3698)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3938..3957)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4285..4306
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4441..4461)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4660..4771
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4889..5029
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4889..5018
                     /label=AAV2 ITR
ORIGIN
        1 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
       61 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
      121 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      181 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
      241 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      301 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
      361 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
      421 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
      481 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      541 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      601 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
      661 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
      721 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      781 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      841 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      901 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
      961 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1021 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1081 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1141 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1201 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1261 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1321 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1381 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1441 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1501 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1561 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1621 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1681 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1741 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2521 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     2581 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2641 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtacca cctgtgtaca
     2701 ttcattggca agtcaccatc ctatcctctt gctgcatctt gcctttccac tgcgattatt
     2761 tttaggggtg tgtgtgcacc gcgctgggaa tatggcaaac gtccccactc cccatcagac
     2821 gtcatgattc ctgcacacac tgggctctca ctaaccccag cttcctttcc cctggtggtt
     2881 ttgagagcat aaattttatt tccagctgtg ggtttgggtc ctatgcagaa cacctccctg
     2941 gagaattgca actaagatgt ttatggcaat cgccctgcaa gtgtctattt aattcctgaa
     3001 aatgtcattt actggtgtgt ttctgtaaaa atcatttctg aaaagttttg tcagcttggc
     3061 ttagcagctg ggggaaaaaa gcagcaaaat gagtcccctc aatcaagcct gtgatttttt
     3121 attttttcag actcatgaat cctcctgaga attaggctta tgacaagcac ccaccccctc
     3181 cacctactcc ctggtttaaa ccaggtgcct gctgacacga aggaagattc cggggcattt
     3241 tgatcctagt cacacagtgt ttctaagtgg gcattactcc tgtcaatcca cagtcttaaa
     3301 gatgttcctc agacaaaaga taattagtga caagaaggcc cactagattc catgtcatat
     3361 cttagttggc gggccagaga aggacagggt ctctggttgg ggccgtgtct cccatcgcat
     3421 tctccctttg caggtgctga agtgaggaag gcccacagct cagccaagca catggggtgc
     3481 ctccctcctg ggccactagc ccactggcca gtaagagaac cagggagcct ttgagctcgg
     3541 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
     3601 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     3661 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3721 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     3781 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     3841 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3901 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3961 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4021 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4081 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     4141 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4201 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     4261 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4321 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     4381 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4441 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4501 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     4561 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4621 caattccgtg gctcgaggac gtgaccgaga gatcttcgac tgtgccttct agttgccagc
     4681 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4741 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4801 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4861 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4921 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4981 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5041 cggtattttc tccttacgca tctgtgcggt atttcacacc gcat
//
