LOCUS       sequence-429272-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SYFP2 in striatal medium spiny neurons in a Cre 
            dependent manner; useful for intersectional labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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: AiP14948:
            pAAV-AiE0441h_3xC2-minBG-DIO-SYFP2-W...
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(825..858)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(909..942)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(960..1670,1671..1673,1674..1676))
                     /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(1002..1023)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1351..1370
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1610..1631
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1673..1682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1699..1732
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1783..1816
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1884..1904)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2091..2202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2320..2460
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2320..2449
                     /label=AAV2 ITR
     rep_origin      2535..2990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2622..2641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2832..2853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3007..3026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3126..3148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3186..3204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3272..3376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3377..3445,3446..4237)
                     /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(3595..3614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4408..4996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4897..4916
                     /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 gcgtcccaca cagcccccac cacctggccg cccacctgaa
      181 aaacttttcc ccaagttctt gagaagtcgc gtaggatcca ggagagaaaa atcaaagtaa
      241 taaaacctca tttaattcct aacggagggc agagctgcga gctgcaatta tcttaatgct
      301 gagctatttt tactctctct ccatctctgt cttctgcctt tctcccacac agcccccacc
      361 acctggccgc ccacctgaaa aacttttccc caagttcttg agaagtcgcg taggatccag
      421 gagagaaaaa tcaaagtaat aaaacctcat ttaattccta acggagggca gagctgcgag
      481 ctgcaattat cttaatgctg agctattttt actctctctc catctctgtc ttctgccttt
      541 ctcccacaca gcccccacca cctggccgcc cacctgaaaa acttttcccc aagttcttga
      601 gaagtcgcgt aggatccagg agagaaaaat caaagtaata aaacctcatt taattcctaa
      661 cggagggcag agctgcgagc tgcaattatc ttaatgctga gctattttta ctctctctcc
      721 atctctgtct tctgcctttc tgagctcggg ctgggcataa aagtcagggc agagccatct
      781 attgcttaca tttgcttctg ggatcctcta gagtcgactc cggaataact tcgtatagga
      841 tactttatac gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa
      901 acctcttaat aacttcgtat agcatacatt atacgaagtt atggcgcgcc gtcgacttac
      961 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     1021 atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg
     1081 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1141 agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc
     1201 ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1261 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1321 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1381 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1441 atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc ccagggtggt cacgagggtg
     1501 ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag
     1561 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     1621 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
     1681 gcgaccggta gcgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta
     1741 acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta
     1801 tgctatacga agttatgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     1861 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1921 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1981 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2041 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     2101 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2161 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2221 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2281 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     2341 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2401 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     2461 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2521 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2581 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2641 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2701 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2761 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2821 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2881 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2941 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     3001 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3061 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3121 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3181 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3241 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3541 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3601 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3901 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4261 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4321 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4381 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4441 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4501 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4561 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4621 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4681 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4741 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4801 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4861 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4921 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4981 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5041 gccttttgct cacatgt
//
