LOCUS       sequence-470331-        4895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0351h is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4895)
  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 4895)
  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: AiP14452:
            pAAV-3xSP10ins-AiE0351h-minBG-SYFP2-...
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   162..302
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   173..302
                     /label=AAV2 ITR
     regulatory      1067..1076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1073..1075,1076..1078,1079..1789)
                     /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(1118..1139)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1379..1398)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1726..1747
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1882..1902)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2101..2212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2330..2459
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2521..3109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2601..2620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3280..4071,4072..4140))
                     /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     3903..3922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4141..4245)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4313..4331
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4369..4391)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4491..4510
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4527..4895,1..87))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4664..4685)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4876..4895
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
       61 cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat
      121 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc
      181 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
      241 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      301 ctgcggccgc acgcgccggt accgaagcta cccctaacac actattctac acacagaaaa
      361 tgctcttcac taggaagcta cccctaacac actattctac acacagaaaa tgctcttcac
      421 taggaagcta cccctaacac actattctac acacagaaaa tgctcttcac tagacgcgta
      481 gactgaggag acgcctcaga ggaatttgta cggggaaagg tctgaggaaa aaggatcgga
      541 tcccaatggg ccgaactcct tcaaaatgtt cccgaggtcc acgtgacgtt ccctcctccc
      601 acccagccct tctttctgtg aagggtgaca cacctcctag aaagaaatct gactcccctc
      661 attgtgtttt taaaataact ctcattattc ctcctctgaa ggttattttc aaactcttta
      721 taactcgatt tttttttaaa tgcaaagaca aggaatcaaa tctgccaact ttctatcaga
      781 gaaactcggg tttctaactt ttaaagtggg attatatgtc ttctccaata tatgggctcc
      841 ctgcttttat aacatgctgc tgttacatct aataaaaacc attttgaatg aaggcagcct
      901 ttcatcgagc tcgattcagc cgggagctta gggaggggag gtcacttcat aagggcttgg
      961 ggggggagtt ggagccacga gtcgtccagc cggagccccg tgtggctgtg ctccggcctc
     1021 agaagcatcc ccggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
     1081 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1141 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1201 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1261 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1321 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1381 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1441 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1501 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1561 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1621 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1681 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1741 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     1801 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     1861 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1921 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1981 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2041 ggctcggctg ttgggcactg acaattccgt ggctcgagca tgcctccgag agatcttcga
     2101 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2161 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2221 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2281 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     2341 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2401 aaggtcgccc gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga
     2461 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2521 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2641 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2701 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2761 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2821 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2881 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2941 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3001 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4261 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4321 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4381 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4441 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4501 gcagattgta ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc
     4561 gttaaatttt tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc
     4621 ttataaatca aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag
     4681 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4741 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     4801 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4861 cgtggcgaga aaggaaggga agaaagcgaa aggag
//
