LOCUS       sequence-472509-        4875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded enhancer AAV vector for multiplexed screening applications;
            AiE0450h is an enhancer active in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4875)
  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 4875)
  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: AiP14450:
            pAAV-AiE0450h-minBG-SYFP2-WPRE3-Barc...
FEATURES             Location/Qualifiers
     source          1..4875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      448..457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(454..456,457..459,460..1170)
                     /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(499..520)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(760..779)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1107..1128
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1263..1283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1482..1593
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1711..1851
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1711..1840
                     /label=AAV2 ITR
     rep_origin      1926..2381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2013..2032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2223..2244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2398..2417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2517..2539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2577..2595)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2663..2767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2768..2836,2837..3628)
                     /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(2986..3005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3799..4387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4288..4307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4449..4578
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ttctaacctg ctggagtgaa gggaaccttg actacttgaa aagcaggctg attgtcaggg
       61 gcggaaccga actcagagct gtcagggccg tcacatggaa agtgcctggc agggggcggc
      121 gacagggggc ttgggctgac agggctgtga ttttgaaaga aacgtctcca agaaagtgat
      181 ttaaagtagg agagaaatat tattattaga taatggctgt caggttattt tttgatgtgt
      241 tttctgagta tttcataatc atcttgcgct tagatggtaa gcccttcctg gctaggggcc
      301 gtgtcccatc ccatgcctct gtggctttca ccttcgggtg tgcaacaggt atcagagctc
      361 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
      421 gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
      481 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      541 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
      601 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
      661 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      721 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      781 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      841 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      901 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
      961 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1021 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1081 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1141 atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt
     1201 cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1261 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1321 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     1381 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1441 gacaattccg tggctcgagt tctgcctcga gagatcttcg actgtgcctt ctagttgcca
     1501 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1561 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1621 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1681 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     1741 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     1801 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     1861 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     1921 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1981 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     2041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2221 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     2281 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     2341 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     2401 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     2461 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     2521 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     2581 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2641 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2701 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2761 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2821 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2881 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2941 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3001 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3061 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3121 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3181 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3241 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3301 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3361 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3421 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3481 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3541 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3601 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4441 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
     4501 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     4561 ccatcactag gggttcctgc ggccgcacgc gtgagaccac aggctgagag gccaggcagg
     4621 gttcaccagg acactctgaa agagctgtct ggctcccagg cccacaggcc ttcgagggaa
     4681 tagacacata agccttgctc ctttttctcc tggtatctgc ccacacccat atcctggaaa
     4741 gctgcgtgat ttatgtccac cagaggggat ttgctctgag gctgtggcct gcaggagcgc
     4801 ccttccgctg gagggtggga ggactgtcta aggcagtcac agcaaggaga ggcacggtgt
     4861 gggcactgct gctat
//
