LOCUS       sequence-472519-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0785m is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  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 4863)
  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: AiP14459:
            pAAV-AiE0785m-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      497..1085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     986..1005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1147..1276
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2009..2018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2015..2017,2018..2020,2021..2731)
                     /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(2060..2081)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2321..2340)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2668..2689
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2824..2844)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3043..3154
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3272..3412
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3272..3401
                     /label=AAV2 ITR
     rep_origin      3487..3942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3574..3593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3784..3805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3959..3978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4078..4100
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4138..4156)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4224..4328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4329..4397,4398..4863,1..326)
                     /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(4547..4566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
       61 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      121 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      181 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      241 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      301 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      361 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      421 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      481 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      601 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1081 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1141 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
     1201 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     1261 atcactaggg gttcctgcgg ccgcacgcgt taaaccttgg actgagccca caggatgctc
     1321 tggaaggacc ttgggaaacc gacacacaaa gcctgtctcg gttttgtttt gttttgtttt
     1381 gtcctccctg tgtcgcatcc acaagtccca agcgtgtagc tcctgggaag ctatgtggct
     1441 gttttcacca aggatgactt ctcagagact gacacccata agagaggccc ctccacccga
     1501 gggtgggagg actctctgtg gcatcaccgg gaggggaggc caggtgtggg cactgcagct
     1561 atttctaatc caccagagga aagagagctt tgactgcttg aaaagcaggc tgattgtcag
     1621 gggcggaacc cagctcagag ctgtcaggga agctcctggc agggggtgga gacagggggc
     1681 ttggactgac aggcctgtga ttttgaaaga aatgtcttta agagagtgac ttaaagtagg
     1741 agagaaatat tattattagg taatgcctgt caggttatat attttttttt gatgtgttct
     1801 tggtatttca taatcatttt gcctttagat ggtaagccct tcctggtcct gggccatgaa
     1861 ctatgcccct cgggtctcac cccagccttg gacatgcaat gggcaccagg aggtagagct
     1921 cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc
     1981 agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
     2041 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2101 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     2161 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
     2221 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2281 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2341 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2401 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2461 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     2521 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     2581 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     2641 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2701 gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat
     2761 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2821 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2881 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2941 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3001 tgacaattcc gtggctcgag cggctaatcg agagatcttc gactgtgcct tctagttgcc
     3061 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3121 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3181 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3241 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3301 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3361 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3421 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3481 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3541 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3601 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3661 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3721 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3781 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3841 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     3901 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3961 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4021 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4081 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4141 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4201 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4261 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4321 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4381 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4441 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4501 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4561 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4621 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4681 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4741 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4801 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4861 cca
//
