LOCUS       sequence-190475-        6060 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of mouse CB2 cannabinoid receptors and eGFP in the 
            presence of Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Li Y, Kim J
  TITLE     Distinct roles of neuronal and microglial CB2 cannabinoid receptors 
            in the mouse hippocampus.
  JOURNAL   Neuroscience. 2017 Sep 6. pii: S0306-4522(17)30629-2. doi: 
            10.1016/j.neuroscience.2017.08.053.
  PUBMED    28888955
REFERENCE   2  (bases 1 to 6060)
  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"; journalName: 
            "Neuroscience. 2017 Sep 6. pii: S0306-4522(17)30629-2. doi: 
            10.1016/j.neuroscience.2017.08.053."
COMMENT     Sequence Label: pAAV-CBh-DIO-CB2-EGFP
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(27..60)
                     /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(111..144)
                     /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(281..994,995..997,998..1000))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(284..994,995..997,998..1000))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(326..347)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     675..694
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     934..955
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1019..1075)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(1085..1108)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2202..2235
                     /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    2286..2319
                     /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(2389..2409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2797..2937
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2797..2926
                     /label=AAV2 ITR
     rep_origin      3012..3467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3099..3118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3309..3330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3484..3503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3603..3625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3663..3681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3749..3853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3854..3922,3923..4714)
                     /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(4072..4091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4885..5473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5374..5393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5535..5664
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        5679..5964
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggatcctc tagagtcgac tccggaataa cttcgtatag gatactttat acgaagttat
       61 gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt
      121 atagcataca ttatacgaag ttatggcgcg ccttatctag atttagccac cttacttgta
      181 cagctcgtcc atgcncgakw rksawymygr mrscksttam taacttcrkm wagcatacat
      241 tatacgaagt tatggcgcgc cttatctaga tttagccacc ttacttgtac agctcgtcca
      301 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
      361 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
      421 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
      481 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
      541 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
      601 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
      661 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
      721 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
      781 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
      841 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
      901 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
      961 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtgccggta gcgctagcag
     1021 gtccagggtt ctcctccacg tctccagcct gcttcagcag gctgaagtta gtagctccgc
     1081 ttcccttgtc atcgtcatcc ttgtaatcct cgagcggccg ggtacgcgtg gtggttttca
     1141 catcagcctc tgtttctgta accgaggacc ttgggccttc ttctttcccc tcaggtccga
     1201 ggccctgtag atacttcttc cagcctatca ggcagtgctg ggcagcagag cgaatctctc
     1261 cactccgcag ggcgtaaatg ataggattga ccatagagtt aacaaggcac agcatggaac
     1321 agaaggcgaa ggcctccttg acctggtcac tcagcgtggt gaccaggctg tggcccatga
     1381 gagccagtgc agggaaccag catatgagca gcacagccag caccaggccc agagtcttgg
     1441 ccaacctcac gtctagccgc atccgagcta tcccaggcac ctgcctgtcc tggtgctcag
     1501 ccaaggtggc tacatgccgg tgggctttcc agaggacata cccataggta tagatgatgc
     1561 cggaaaagag gatggcaatg aatagaagcc agcccagtag gtagtcgtta gggatcagtg
     1621 ggaaaagctc agagcaggga ctagggcaac aagtccaccc catgagcggc aggtaagaaa
     1681 tcaatgctga gaggacccac atgacacaga gggccaccag tgccctccca cgggtgacta
     1741 gagctttgta ggtaggcggg taacacagac ataggtagcg gtcaacagcg gttagcagca
     1801 ggctgcccac agaggctgtg aaggtcatgg tcacactgcc gatcttcagc aggaagatag
     1861 cgttggagtc gaccccgtgg aagacgtgga agatgacaaa gttgcaggcg aagatcacgc
     1921 tggccaggaa gtcagctcca gccaagctgc tgatgaacag gtacgagggc tttctgcgga
     1981 gccgccggga ggacaggata atatagagca cggccatgtt ctccagggcg ctcagcagcc
     2041 ccatcagggt gcacagcacc gccacggcga tctgctggcc actgctcagg atcatgtact
     2101 ccttcatggg gttgaactcc aagccaccgt tggagccgtt ggtcacttct gtctcccggc
     2161 atccctccat ggcgatcgcg gcggcagatc tcctcgctag cataacttcg tataaagtat
     2221 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     2281 aaagaataac ttcgtataat gtatgctata cgaagttatg aattccgctc gagataatca
     2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2461 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2521 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2581 gtttatttgt gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg
     2641 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2701 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     2761 gcggccgcgg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2821 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2881 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2941 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3001 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3061 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3361 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3481 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3661 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3841 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3901 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3961 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4021 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4081 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4141 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4201 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4261 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4321 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4381 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4441 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4501 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4561 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4621 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4681 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4741 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4801 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4861 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5521 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     5581 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     5641 ccaactccat cactaggggt tcctgcggcc gcacgcgtcg ttacataact tacggtaaat
     5701 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     5761 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5821 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5881 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     5941 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     6001 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
//
