LOCUS       sequence-385260-        7392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of mEGFP-DVL1 WT Fusion Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  AUTHORS   Mensah MA, Niskanen H, Magalhaes AP, Basu S, Kircher M, Sczakiel HL,
            Reiter AMV, Elsner J, Meinecke P, Biskup S, Chung BHY, Dombrowsky G,
            Eckmann-Scholz C, Hitz MP, Hoischen A, Holterhus PM, Hulsemann W, 
            Kahrizi K, Kalscheuer VM, Kan A, Krumbiegel M, Kurth I, Leubner J, 
            Longardt AC, Moritz JD, Najmabadi H, Skipalova K, Snijders Blok L, 
            Tzschach A, Wiedersberg E, Zenker M, Garcia-Cabau C, Buschow R, 
            Salvatella X, Kraushar ML, Mundlos S, Caliebe A, Spielmann M, Horn 
            D, Hnisz D
  TITLE     Aberrant phase separation and nucleolar dysfunction in rare genetic 
            diseases.
  JOURNAL   Nature. 2023 Feb;614(7948):564-571. doi: 10.1038/s41586-022-05682-1.
            Epub 2023 Feb 8.
  PUBMED    36755093
REFERENCE   2  (bases 1 to 7392)
  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/s41586-022-05682-1"; journalName: "Nature"; date: 
            "2023-02"; volume: "614"; issue: "7948"; pages: "564-571"
COMMENT     Sequence Label: pRK5-mEGFP-DVL1-WT
FEATURES             Location/Qualifiers
     source          1..7392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      933..942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(939..941,942..944,945..1652)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(984..1005)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1592..1613
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3692..3701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3705..3839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3742..3761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3796..3815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3908..4265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3908..4099
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3931..3951)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4116..4251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4285..4302)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4285..4301)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4294..4316)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4514..4969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4601..4620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4811..4832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4986..5005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5105..5127
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5165..5183)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5251..5355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5356..5424,5425..6216)
                     /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(5574..5593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6387..6975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6876..6895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7129..7146
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7263..7284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7299..7304,7305..7322,7323..7329)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7337..7353
                     /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)."
     primer_bind     7342..7364
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7361..7377
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7361..7377
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtaccg gtgccaccat ggtgagcaag ggcgaggagc
      961 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1021 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     1081 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1141 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1201 ccatgcccga aggctacgtc caggagcgca ccatctcctt caaggacgac ggcacctaca
     1261 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1321 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     1381 gccacaacgt ctatatcacg gccgacaagc agaagaacgg catcaaggcg aacttcaaga
     1441 tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1501 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     1561 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1621 ccgggatcac tctcggcatg gacgagctgt acacggcgat ggcggagacc aagattatct
     1681 accacatgga cgaggaggag acgccgtacc tggtcaagct gcccgtggcc cccgagcgcg
     1741 tcacgctggc cgacttcaag aacgtgctca gcaaccggcc cgtgcacgcc tacaaattct
     1801 tctttaagtc catggaccag gacttcgggg tggtgaagga ggagatcttt gatgacaatg
     1861 ccaagcttcc ctgcttcaac ggccgcgtgg tctcctggct ggtcctggct gagggtgctc
     1921 actcggatgc ggggtcccag ggcacggaca gccacacaga cctgcccccg cctcttgagc
     1981 ggacaggcgg catcggggac tcccggcccc cctccttcca cccaaatgtg gccagcagcc
     2041 gtgacgggat ggacaacgag acaggcacgg agtccatggt cagtcaccgg cgggagcgtg
     2101 cccgacgccg gaaccgcgag gaggccgccc ggaccaatgg gcacccaagg ggagaccgac
     2161 ggcgggatgt ggggctgccc ccagacagcg cgtccaccgc cctcagcagc gagcttgagt
     2221 ccagcagctt tgtggactcg gacgaggatg gcagcacgag caggctcagc agctccacgg
     2281 agcagagcac ctcatccaga ctcatccgga agcacaaacg ccggcggagg aagcagcgcc
     2341 ttcggcaggc ggaccgggcc tcctccttca gcagcataac cgactccacc atgtccctca
     2401 acatcgtcac tgtcacgctc aacatggaaa gacatcactt tctgggcatc agcatcgtgg
     2461 ggcagagcaa cgaccgtgga gacggcggca tctacattgg ctccatcatg aagggcgggg
     2521 ctgtggccgc tgacggccgc atcgagcccg gcgacatgtt gctgcaggtg aatgacgtga
     2581 actttgagaa catgagcaat gacgatgccg tgcgggtgct gcgggagatc gtttcccaga
     2641 cggggcccat cagcctcact gtggccaagt gctgggaccc aacgccccga agctacttca
     2701 ccgtcccacg ggctgacccg gtgcggccca tcgaccccgc cgcctggctg tcccacacgg
     2761 cggcactgac aggagccctg ccccgctacg agctggaaga ggcgccgctg acggtgaaga
     2821 gtgacatgag cgccgtcgtc cgggtcatgc agctgcccga ttctgggctc gaaattaggg
     2881 ataggatgtg gctgaaaatt actatagcta acgcagtaat tggcgccgat gtcgttgatt
     2941 ggttatatac ccatgtcgaa gggtttaaag aaagacgtga agcgaggaaa tatgcttctt
     3001 ctctcttgaa acatgggttt cttcgccata ccgtgaataa aatcactttc tcagaacaat
     3061 gttattatgt gttcggagac ttgtgtagta acttggctac attaaatctg aattcagggt
     3121 catctggaac atccgaccaa gataccctcg ctcccttgcc acatcctgcc gcaccatggc
     3181 ccctcgggca aggttatcca tatcaatatc cagggccgcc gccatgtttt cccccggctt
     3241 atcaagatcc tggtttctcc tacggttctg gttcaacggg atctcaacaa tcagagggct
     3301 ccaagtccag cggatctact cgctcttcac gaagggcacc tggcagggaa aaggaacgcc
     3361 gagccgcagg cgctggcggt agcggatccg agtctgacca tacagctcca tctggcgtag
     3421 gatcctcatg gagagaacgg cccgcagggc aactgtctag aggatccagt cctagaagcc
     3481 aagcgagcgc gactgcaccc ggccttccac cacctcatcc tacaactaaa gcttacaccg
     3541 ttgtcggtgg tccgccaggc ggcccgcccg tgagagaact cgcagctgtg ccacccgagt
     3601 taaccggatc aagacaaagc tttcaaaagg ctatggggaa cccctgcgag ttcttcgtgg
     3661 acatcatgtg aagtcgacct gcagaagctt ggccgccatg gcccaacttg tttattgcag
     3721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     3841 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     3901 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4141 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4201 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4261 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4321 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4381 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4441 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4501 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4561 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4621 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4681 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4741 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4801 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     4861 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4921 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4981 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5041 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5101 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5161 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5221 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5281 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5341 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5401 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5461 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5521 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5581 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5641 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5701 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5761 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5821 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5881 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5941 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6001 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6061 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6121 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6181 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6241 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6301 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6361 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6421 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6481 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6541 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6601 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6661 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6721 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6781 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6841 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6901 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6961 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7021 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7081 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7141 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7201 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7261 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7321 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
     7381 attacgaatt aa
//
