LOCUS       sequence-472516-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0367h is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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: AiP14448:
            pAAV-3xSP10ins-AiE0367h-minRho-SYFP2...
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     212..233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(387..406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     506..528
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(566..584)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        652..756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(757..825,826..1617)
                     /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(975..994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1788..2376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2277..2296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2438..2567
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3310..3319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3316..3318,3319..3321,3322..4032)
                     /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(3361..3382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3622..3641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3969..3990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4125..4145)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4344..4455
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4573..4713
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4573..4702
                     /label=AAV2 ITR
     rep_origin      join(4788..4873,1..370)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
       61 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      121 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
      241 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
      301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
      361 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      421 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
      481 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
      541 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      601 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      661 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      721 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      781 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      841 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      901 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      961 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1021 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1081 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1141 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1201 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1261 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1321 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1381 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1441 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     1501 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1561 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1621 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1681 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1741 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1801 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1861 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1921 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1981 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2041 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2101 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2161 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2221 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2281 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2341 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2401 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
     2461 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     2521 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg
     2581 ccggtaccga agctacccct aacacactat tctacacaca gaaaatgctc ttcactagga
     2641 agctacccct aacacactat tctacacaca gaaaatgctc ttcactagga agctacccct
     2701 aacacactat tctacacaca gaaaatgctc ttcactagac gcgttagcag gagtacatcc
     2761 tcctctgtta gaaaaacaac tctgcggttc tgggtcactt ttcccatctt tgtatagaaa
     2821 agtaccgatt attattgttc ccccgggtgc cttctccagc ttaaaggaaa attccagccc
     2881 cagagatctt ccgcagtgag tgacagctcc ttaatgtgag gagcagcgtg ggcgtggtgg
     2941 gtgggatgtc agagccatca aagccctcat ttctgatccc tctgtgatgt cggggttccc
     3001 tggcacgccc actgtcatgc ctcagcacat tggcatctga gagaggaaag gtgcagctga
     3061 caccctgacc cccttctccc cgacacgggg tcagacaggg tcccgcggtg gcctggttgt
     3121 gcagcactga catgttgcct gggaacgaag agctcgattc agccgggagc ttagggaggg
     3181 gaggtcactt cataagggct tgggggggga gttggagcca cgagtcgtcc agccggagcc
     3241 ccgtgtggct gtgctccggc ctcagaagca tccccggatc cagatctttc gaagctagcg
     3301 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3361 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3421 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     3481 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     3541 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3601 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3661 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3721 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     3781 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     3841 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3901 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     3961 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4021 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     4081 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4141 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4201 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     4261 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     4321 gtgctatatc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4381 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4441 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4501 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     4561 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4621 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4681 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4741 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4801 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4861 cgccttagcg ccc
//
