LOCUS       sequence-472836-        4708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0441h is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4708)
  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 4708)
  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: AiP14453:
            pAAV-AiE0441h-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..4708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   88..217
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(279..867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(359..378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1038..1829,1830..1898))
                     /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     1661..1680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1899..2003)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2071..2089
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2127..2149)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2249..2268
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2285..2740)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2422..2443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2634..2653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2815..2955
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2826..2955
                     /label=AAV2 ITR
     polyA_signal    3073..3184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3383..3403
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3496..4206,4207..4209,4210..4212))
                     /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(3538..3559)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3887..3906
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4146..4167
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4209..4218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gggactcctg agagctggag gaggggtggg caggggcaag gagtggggag gggctgctgg
       61 ggacagtgga gtcacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
      121 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
      181 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
      241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2041 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2101 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2161 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2221 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2281 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2341 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     2401 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2461 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2521 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2581 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2641 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2701 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     2761 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     2821 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     2881 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     2941 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     3001 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3061 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3121 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3181 acagtcgaag atctctcgat ctctacctcg agccacggaa ttgtcagtgc ccaacagccg
     3241 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa
     3301 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     3361 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     3421 caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg
     3481 cgcgccgtcg acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
     3541 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
     3601 gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     3661 ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt
     3721 ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt
     3781 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     3841 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     3901 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     3961 gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag
     4021 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag
     4081 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     4141 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     4201 cttgctcacc atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca
     4261 aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct cgtggggaag
     4321 actggctgac agagggacag acatgaagga ggaggaagag actgcacaga gacaggacgt
     4381 gagagagaga ttggggggag agaggagaaa gcaaagagag acagaaaaca tggttgagag
     4441 atggagagac aaagggaggc agagagagag acagcgaacc tcacaacaga gaaaggcaga
     4501 agacagagat ggagagagag taaaaatagc tcagcattaa gataattgca gctcgcagct
     4561 ctgccctccg ttaggaatta aatgaggttt tattactttg atttttctct cctggatcct
     4621 acgcgacttc tcaagaactt ggggaaaagt ttttcaggtg ggcggccagg tggtgggggc
     4681 tgtgtgggcg acagaggagg aagggaat
//
