LOCUS       sequence-244372-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  labelling of neuronal nuclear envelope with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  AUTHORS   Obata Y, Castano A, Boeing S, Bon-Frauches AC, Fung C, Fallesen T, 
            de Aguero MG, Yilmaz B, Lopes R, Huseynova A, Horswell S, Maradana 
            MR, Boesmans W, Vanden Berghe P, Murray AJ, Stockinger B, Macpherson
            AJ, Pachnis V
  TITLE     Neuronal programming by microbiota regulates intestinal physiology.
  JOURNAL   Nature. 2020 Feb;578(7794):284-289. doi: 10.1038/s41586-020-1975-8. 
            Epub 2020 Feb 5.
  PUBMED    32025031
REFERENCE   2  (bases 1 to 6345)
  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: 
            "10.1038/s41586-020-1975-8"; journalName: "Nature"; date: "2020-02";
            volume: "578"; issue: "7794"; pages: "284-289"
COMMENT     Sequence Label: pAAV-CaMKII-EGFPKASH
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        154..1442
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1463..1472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1469..1471,1472..1474,1475..2185)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1469..1471,1472..1474,1475..2185)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1514..1535)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1775..1794)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2122..2143
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2477..3065
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2530..2550)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2948..2959)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3097..3573
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3613..3753
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3613..3742
                     /label=AAV2 ITR
     rep_origin      3828..4283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3915..3934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4125..4146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4300..4319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4419..4441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4479..4497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4565..4669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4670..4738,4739..5530)
                     /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(4888..4907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5701..6289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6190..6209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt taacattatg gccttaggtc acttcatctc
      181 catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag
      241 tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat
      301 cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag
      361 ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc
      421 gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct
      481 accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat
      541 ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct
      601 gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg
      661 tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag
      721 cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc
      781 cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt
      841 agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct
      901 cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca
      961 gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat
     1021 cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac
     1081 ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg
     1141 aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag
     1201 ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg
     1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg
     1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag
     1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca
     1441 gctctagagc ggtaccggat ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1501 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1561 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1621 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     1681 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1741 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1801 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1861 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1921 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1981 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2041 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2101 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2161 tctcggcatg gacgagctgt acaagtccgg actcagatct cgagaggagg aggaggagac
     2221 agacagcagg atgccccacc tcgacagccc cggcagctcc cagccgagac gctccttcct
     2281 ctcaagggtg atcagggcag cgctaccgtt gcagctgctt ctgctgctgc tgctgctcct
     2341 ggcctgcctg ctacctgcct ctgaagatga ctacagctgc acccaggcca acaactttgc
     2401 ccgatccttc taccccatgc tgcggtacac caacgggcca cctcccacct aggaattcga
     2461 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2521 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2581 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2641 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2701 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccggaacttt
     2761 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2821 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2881 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2941 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3001 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3061 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     3121 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3181 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     3241 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     3301 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     3361 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     3421 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     3481 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     3541 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     3601 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3661 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3721 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3781 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4201 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4261 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4321 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4381 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4441 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4501 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5521 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctca
//
