LOCUS       sequence-354794-        4033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Notch reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4033)
  AUTHORS   Hansson EM, Teixeira AI, Gustafsson MV, Dohda T, Chapman G, Meletis 
            K, Muhr J, Lendahl U
  TITLE     Recording Notch signaling in real time.
  JOURNAL   Dev Neurosci. 2006;28(1-2):118-27.
  PUBMED    16508309
REFERENCE   2  (bases 1 to 4033)
  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: "Dev 
            Neurosci."; date: "2006"; volume: "28(1-2)"; pages: "118-27"
COMMENT     Sequence Label: 12XCSL-d1EGFP
FEATURES             Location/Qualifiers
     source          1..4033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..98
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     90..107
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     91..107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    533..585
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(599..601,602..604,605..1315,1316..1321,1322..1444)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to a mutated version 
                     of residues 422-461 of mouse ornithine decarboxylase, 
                     giving an in vivo half-life of ~1 hour"
                     /label=d1EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1252..1273
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1566..1687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1603..1622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1657..1676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1716..1732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1716..1732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1729..1751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1740..1756
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1764..1770,1771..1788,1789..1794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1809..1830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1947..1964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2118..2706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2198..2217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2877..3668,3669..3737))
                     /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     3500..3519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3738..3842)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3910..3928
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
       61 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat
      121 gcctgcaggt cgactctaga ggatcccgac tcatgggaaa atgggcggaa gggcaccgtg
      181 ggaaaatagt agatcccgac tcgtgggaaa atgggcggaa gggcaccgtg ggaaaatagt
      241 agatcccgac tcgtgggaaa atgggcggaa gggcaccgtg ggaaaatagt agatcccgac
      301 tcatgggaaa atgggcggaa gggcaccgtg ggaaaatagt agatcccgac tcgtgggaaa
      361 atgggcggaa gggcaccgtg ggaaaatagt agatcccgac tcgtgggaaa atgggcggaa
      421 gggcaccgtg ggaaaatagt agatccactg tatgcatctt gggcataaaa ggcagatcac
      481 ttcagcttct gcttacactt gcttttgaca caactgtgtt tacttgcaat ccctcgagct
      541 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      721 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1081 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagaagct
     1321 tagccatggc ttcccgccgg cggtggcggc gcaggatgat ggcacgctgc ccatgtcttg
     1381 tgcccaggag agcgggatgg accgtcaccc tgcagcctgt gcttctgcta ggatcaatgt
     1441 gtaggcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     1501 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1561 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1621 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1681 tatcttaagg gtaccgagct cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     1741 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3841 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3901 acctataaaa ataggcgtat cacgaggcct atgcggtgtg aaataccgca cagatgcgta
     3961 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4021 cgatcggtgc ggg
//
