LOCUS       sequence-425145-        5390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SYFP2 (yellow fluorescent protein) in striatal direct 
            pathway medium spiny neurons (D1 MSNs), stronger than 3xCore version
            CN3044).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  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 5390)
  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: AiP14696: pAAV-AiE0779m_6xC2-minBG-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5390
                     /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      1402..1411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1408..1410,1411..1413,1414..2124)
                     /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(1453..1474)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1714..1733)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2061..2082
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2217..2237)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2424..2535
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2653..2793
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2653..2782
                     /label=AAV2 ITR
     rep_origin      2868..3323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2955..2974)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3165..3186
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3340..3359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3459..3481
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3519..3537)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3605..3709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3710..3778,3779..4570)
                     /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(3928..3947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4741..5329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5230..5249
                     /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 tcaagtttga gacaaattaa ggaaaagcgg gttgggagga gtctgaggct
      781 gtgggtggct gcctcgagag gctgctcaca acctgacatg tttcagatct tttgacacgg
      841 tcatgcaagc aacagaaact tatatggttt ttagtctccc aggtagcagg gaaatgatga
      901 tgtgggagga ttaggggggg gaaaataagt ttgagacaaa ttaaggaaaa gcgggttggg
      961 aggagtctga ggctgtgggt ggctgcctcg agaggctgct cacaacctga catgtttcag
     1021 atcttttgac acggtcatgc aagcaacaga aacttatatg gtttttagtc tcccaggtag
     1081 cagggaaatg atgatgtggg aggattaggg gggggaaaat aagtttgaga caaattaagg
     1141 aaaagcgggt tgggaggagt ctgaggctgt gggtggctgc ctcgagaggc tgctcacaac
     1201 ctgacatgtt tcagatcttt tgacacggtc atgcaagcaa cagaaactta tatggttttt
     1261 agtctcccag gtagcaggga aatgatgatg tgggaggatt agggggggga aaatgggctg
     1321 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     1381 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1441 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1501 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1561 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1621 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1681 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1741 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1801 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1861 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1921 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1981 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     2041 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2101 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     2161 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2221 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2281 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2341 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2401 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2461 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2521 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2581 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2641 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2701 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2761 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2821 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2881 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2941 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3001 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3061 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3121 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3181 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3241 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3301 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3421 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3481 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3541 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3601 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3661 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3721 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3781 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3841 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3901 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3961 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4021 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4081 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4141 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4201 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4261 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4321 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4381 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4441 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4501 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4561 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
