LOCUS       sequence-379462-        4808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AI plasmid ALIAS: AiP13044. Optimized expression of SYFP2 (yellow 
            fluorescent protein) in striatal direct pathway medium spiny neurons
            (D1-MSNs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4808)
  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 4808)
  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: AiP13044 -
            pAAV-AiE0779m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      820..829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(826..828,829..831,832..1542)
                     /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(871..892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1132..1151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1479..1500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1635..1655)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1842..1953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2071..2211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2071..2200
                     /label=AAV2 ITR
     rep_origin      2286..2741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2373..2392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2583..2604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2758..2777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2877..2899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2937..2955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3023..3127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3128..3196,3197..3988)
                     /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(3346..3365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4159..4747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4648..4667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagttt gagacaaatt aaggaaaagc gggttgggag
      181 gagtctgagg ctgtgggtgg ctgcctcgag aggctgctca caacctgaca tgtttcagat
      241 cttttgacac ggtcatgcaa gcaacagaaa cttatatggt ttttagtctc ccaggtagca
      301 gggaaatgat gatgtgggag gattaggggg gggaaaataa gtttgagaca aattaaggaa
      361 aagcgggttg ggaggagtct gaggctgtgg gtggctgcct cgagaggctg ctcacaacct
      421 gacatgtttc agatcttttg acacggtcat gcaagcaaca gaaacttata tggtttttag
      481 tctcccaggt agcagggaaa tgatgatgtg ggaggattag gggggggaaa ataagtttga
      541 gacaaattaa ggaaaagcgg gttgggagga gtctgaggct gtgggtggct gcctcgagag
      601 gctgctcaca acctgacatg tttcagatct tttgacacgg tcatgcaagc aacagaaact
      661 tatatggttt ttagtctccc aggtagcagg gaaatgatga tgtgggagga ttaggggggg
      721 gaaaatgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      781 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
      841 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      901 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      961 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1021 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1081 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1141 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1201 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1261 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1321 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1441 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1501 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg
     1561 cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact
     1621 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1681 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     1741 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1801 ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca
     1861 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1921 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1981 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2041 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2101 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2161 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     2221 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     2281 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2341 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     2401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2581 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     2641 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     2701 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     2761 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     2821 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     2881 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     2941 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3001 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3061 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3121 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3181 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3241 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3301 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3361 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3421 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3481 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3541 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3601 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3661 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3721 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3781 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3841 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3901 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3961 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4021 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4801 tcacatgt
//
