LOCUS       sequence-470329-        4895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0783m 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: AiP14454:
            pAAV-AiE0783m-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(50..118,119..910)
                     /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(268..287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1081..1669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1570..1589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1731..1860
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2647..2656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2653..2655,2656..2658,2659..3369)
                     /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(2698..2719)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2959..2978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3306..3327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3462..3482)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3681..3792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4103..4558
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4190..4209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4400..4421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4575..4594)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4694..4716
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4754..4772)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4840..4895,1..49)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
       61 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      121 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      181 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      241 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      301 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      361 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      421 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      481 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      541 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
      601 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
      661 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
      721 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
      781 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
      841 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
      901 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
      961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1081 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1141 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1201 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1261 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1321 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1381 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1441 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1501 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1561 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1621 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1681 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     1741 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     1801 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     1861 gcggccgcac gcgttccttg gacactgaat tctgagagac agaggtagga gaggggaacc
     1921 atgggatgtg acgatttcaa agatttaccc tgggcactgt ataagaaaac gttttccaaa
     1981 gtgtctgaca ctgggtctga ctgacacaag gcaccagcaa atgtcacctc tcttccctgt
     2041 gtgtcacaca acttcatttc tcttgaaact aaaacttctc ttctagttgc ccttgacttc
     2101 caatgtcacc gttctctagc ccatacccca ttactaatcc tccccgcccg gttccgccta
     2161 cgtccctttt gttcccacta aagatggctt tgctttttca cctgtttaag tcttcctgcc
     2221 tcccacactg atgaattcag ttagactgaa aaaaaaaaaa agaggcctct tcctccataa
     2281 aaccccaatt aatcaatcat tcacgactga gcaatttggg gtgatggtat ttagtgggaa
     2341 gaactgctcc agcactgtca ggcctgggat gtgacactaa cagcagtgag acctaagagg
     2401 agtaatttgg cttttttcat atctcagttt gctcactcac agtaccttaa ctcttaagac
     2461 cggtaagagt gaaaattacc cccagaggaa tcctagggca aagacagcca ggtgatcatc
     2521 agtataaact aaggatgcct gttcatacca gcagagctcg ggctgggcat aaaagtcagg
     2581 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
     2641 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2701 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2761 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     2821 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     2881 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2941 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3001 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3061 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3121 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     3181 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3241 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     3301 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3361 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     3421 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3481 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3541 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     3601 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagaa
     3661 ctgtagcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3721 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3781 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3841 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     3901 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3961 tcactgaggc cgggcgacca aaggtcgccc ggcctcagtg agcgagcgag cgcgcagctg
     4021 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4081 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4141 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4201 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4261 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4321 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4381 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4441 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4501 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4561 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4621 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4681 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4741 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4801 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4861 tttttctaaa tacattcaaa tatgtatccg ctcat
//
