LOCUS       sequence-238517-        6356 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 6356)
  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 6356)
  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..6356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1469..1478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1475..1477,1478..1480,1481..2191)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1475..1477,1478..1480,1481..2191)
                     /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(1520..1541)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1781..1800)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2128..2149
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2483..3071
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2536..2556)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2954..2965)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3103..3579
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3302..3321)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3619..3759
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3619..3748
                     /label=AAV2 ITR
     rep_origin      3834..4289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3921..3940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4131..4152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4306..4325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4425..4447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4485..4503)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4571..4675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4676..4744,4745..5536)
                     /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(4894..4913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5707..6295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6196..6215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatccgc caccatggtg agcaagggcg aggagctgtt
     1501 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1561 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1621 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1681 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1741 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1801 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1861 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1921 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1981 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2041 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2101 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2161 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag aggaggagga
     2221 ggagacagac agcaggatgc cccacctcga cagccccggc agctcccagc cgagacgctc
     2281 cttcctctca agggtgatca gggcagcgct accgttgcag ctgcttctgc tgctgctgct
     2341 gctcctggcc tgcctgctac ctgcctctga agatgactac agctgcaccc aggccaacaa
     2401 ctttgcccga tccttctacc ccatgctgcg gtacaccaac gggccacctc ccacctagga
     2461 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2521 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2581 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2641 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2701 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2761 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2821 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2881 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2941 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3001 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3061 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3121 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3181 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3241 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3301 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3361 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3421 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3481 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3541 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3601 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3661 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3721 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3781 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3961 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4021 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4141 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4201 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4261 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4321 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4381 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4441 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4501 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4561 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4621 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4681 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4741 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4801 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4861 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4921 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4981 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5041 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5101 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5161 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5221 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5281 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5341 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5401 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5461 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5521 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5581 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5641 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5701 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5761 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5821 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5881 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5941 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6001 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6061 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
