LOCUS       sequence-484259-        4959 bp    DNA     linear   UNA 09-OCT-2025
DEFINITION  Cre dependent flag-tagged Rpl22l1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4959)
  AUTHORS   Tanimura A, Du Y, Kondapalli J, Wokosin DL, Surmeier DJ
  TITLE     Cholinergic Interneurons Amplify Thalamostriatal Excitation of 
            Striatal Indirect Pathway Neurons in Parkinson's Disease Models.
  JOURNAL   Neuron. 2019 Feb 6;101(3):444-458.e6. doi: 
            10.1016/j.neuron.2018.12.004. Epub 2019 Jan 15.
  PUBMED    30658860
REFERENCE   2  (bases 1 to 4959)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2018.12.004"; journalName: "Neuron"; date: 
            "2019-02-6- 6"; volume: "101"; issue: "3"; pages: "444-458.e6"
COMMENT     Sequence Label: pFB-EF1a-DIO-Rpl22l1-myc-Flag-2A-tdTomato-WPRE...
FEATURES             Location/Qualifiers
     source          1..4959
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1450..1483)
                     /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(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1576..3006)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             complement(3007..3060)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(3076..3099)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(3118..3147)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3525..3534
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3541..3574
                     /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    3625..3658
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3683..4271
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3736..3756)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4154..4165)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4303..4779
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4502..4521)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4819..4959
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4819..4948
                     /label=AAV2 ITR
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttact tgtacagctc gtccatgccg tacaggaaca ggtggtggcg
     1621 gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc
     1681 cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg ccatgtagat
     1741 ggtcttgaac tccaccaggt agcggccgcc gtccttcagc ttcagggcct ggtggatctc
     1801 gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc agcccatggt
     1861 cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta
     1921 gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc
     1981 gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct tgtaatcggg
     2041 gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg gggacaggat
     2101 gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc
     2161 ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcatggagcc
     2221 ctccatgcgc accttgaagc gcatgaactc tttgatgacg gccatgttgt tgtcctcgga
     2281 ggaggcggtg ccggagctgc cgctgccggt gctgccggtg ccatgcccca ggaacaggtg
     2341 gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     2401 gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat
     2461 gtagatggtc ttgaactcca ccaggtagcg gccgccgtcc ttcagcttca gggcctggtg
     2521 gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc
     2581 catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac
     2641 cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc
     2701 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta
     2761 atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga
     2821 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg
     2881 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat
     2941 ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct cgcccttgct
     3001 caccattggg ccaggattct cctcgacgtc accgcatgtt agcagacttc ctctgccctc
     3061 tccactgcca gaaaccttat cgtcgtcatc cttgtaatcc aggatatcat ttgctgccag
     3121 atcctcttct gagatgagtt tctgctcgag cggccgcgta cgcgtgtcct cagactctga
     3181 accatcttca tcttgactaa tctggaaata acgaagttcg taagtctcct tgtcagatgc
     3241 gaccacacgt agccaatcac ggagattatt cttcttaagg tatttcttgg taagatattt
     3301 caaatacctt ttggagaact gtttctcaga aacaactgtg attttattct tcaggcgttc
     3361 aatgtgaaca acattgccaa gatttccagt ttttccattg actttaacct tctcccgtag
     3421 aaactgttca aaatttccag agtcaaaaat tccatcttct actggatgag taaggtccaa
     3481 atgaaacctc caggtggact tcttaggctt cttgtctttc ggcgccatgg tggcgctagc
     3541 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3601 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3661 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3721 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3781 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3841 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3901 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3961 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4021 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4081 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4141 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4201 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4261 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     4321 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4381 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4441 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4501 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4561 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4621 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4681 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4741 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4801 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4861 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4921 cggcctcagt gagcgagcga gcgcgcagct gcctgcagg
//
