LOCUS       sequence-406608-        7857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, CDK6_SFTA3 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  AUTHORS   Tripathi S, Shirnekhi HK, Gorman SD, Chandra B, Baggett DW, Park CG,
            Somjee R, Lang B, Hosseini SMH, Pioso BJ, Li Y, Iacobucci I, Gao Q, 
            Edmonson MN, Rice SV, Zhou X, Bollinger J, Mitrea DM, White MR, 
            McGrail DJ, Jarosz DF, Yi SS, Babu MM, Mullighan CG, Zhang J, Sahni 
            N, Kriwacki RW
  TITLE     Defining the condensate landscape of fusion oncoproteins.
  JOURNAL   Nat Commun. 2023 Sep 28;14(1):6008. doi: 10.1038/s41467-023-41655-2.
  PUBMED    37770423
REFERENCE   2  (bases 1 to 7857)
  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: "Nat 
            Commun."; date: "2023-09-28"; volume: "14(1):6008. doi"; pages: " 
            10.1038/s41467-023-41655-2"
COMMENT     Sequence Label: CL20_mEGFP_CDK6_SFTA3
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     433..452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(671..775)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        806..1185
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             1187..1444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1491..1616
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2108..2225
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2585..2818
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3003..3047
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        3304..3607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3608..3811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3762..3782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3887..3896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3893..3895,3896..3898,3899..4609)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3938..3959)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4199..4218)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4546..4567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5680..5699)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5745..5800
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5799..5818)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6161..6177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6161..6177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6174..6196)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6185..6201
                     /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(6209..6215,6216..6233,6234..6239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6254..6275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6334..6529
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6380..6515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6535..6669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6572..6591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6626..6645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6737..6754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6908..7496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6988..7007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7667..7857,1..601,602..670))
                     /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"
ORIGIN
        1 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
       61 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
      121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
      181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
      241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
      301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      781 ttccccgaaa agtgccacct gggtcgacat tgattattga ctagttatta atagtaatca
      841 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      901 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      961 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1021 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1081 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1141 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggagg cgtggcctgg
     1201 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     1261 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     1321 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     1381 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1441 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1501 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1561 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1621 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1681 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1741 atactggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1801 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1861 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1921 agcaaaacaa aagtaagaaa aaagcacagc aagcagcagg atcttcagac ctggaaattc
     1981 cctacaatcc ccaaagtcaa ggagtagtag aatctatgaa taaagaatta aagaaaatta
     2041 taggacaggt aagagatcag gctgaacatc ttaagacagc agtacaaatg gcagtattca
     2101 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2161 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2221 attttcgggt ttattacagg gacagcagaa atccactttg gaaaggacca gcaaagctcc
     2281 tctggaaagg tgaaggggca gtagtaatac aagataatag tgacataaaa gtagtgccaa
     2341 gaagaaaagc aaagatcatt agggattatg gaaaacagat ggcaggtgat gattgtgtgg
     2401 caagtagaca ggatgaggat tagaacatgg aaaagtttag taaaacacca taaggaggag
     2461 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     2521 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     2581 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     2641 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     2701 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     2761 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     2821 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     2881 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2941 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     3001 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     3061 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     3121 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     3181 cattatcgtt tcagacccac ctcccaaccc cgaggggacc gagctcaagc ttcgaacgcg
     3241 tgctagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     3301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     3361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     3421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     3481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     3541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     3601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     3661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     3721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     3781 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgggctc gtttagtgaa
     3841 ccgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag
     3901 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3961 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4021 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4081 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4141 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4201 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4261 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4321 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4381 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4441 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4501 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4561 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggaagcgg
     4621 aggctccggg ggaagtgcgg ccgcttcccg gatggagaaa gacggattat gccgtgccga
     4681 ccaacaatac gaatgcgtgg ctgagatcgg ggaaggcgcc tatggcaaag tatttaaagc
     4741 acgtgacttg aagaacggcg gacgttttgt agcattaaag cgcgttcgtg ttcaaactgg
     4801 cgaagaaggt atgccactgt caacgatccg cgaggtggcc gtactgcgcc acctggagac
     4861 gtttgaacat ccaaacgtgg tgcgcctttt cgacgtttgc acggtgtctc gcaccgatcg
     4921 cgagacgaaa cttacgttgg tgtttgaaca tgtcgatcaa gatttgacga catacttgga
     4981 taaggttcct gaacccggcg tccccactga aactatcaaa gatatgatgt tccagttatt
     5041 acgtggcctt gacttcttac attcgcaccg tgtagtacat cgtgatttaa aaccccagaa
     5101 catcttagtc acatcatccg gccagatcaa gttagccgat tttggacttg cccgtattta
     5161 tagttttcaa atggccctta cgtcagtggt tcttttttta caagatcagg cacagcgcct
     5221 taccgaatgg ctgcagttat caggttttga gaatcctgtc tctgagagta cgacattatg
     5281 cttacgtgaa cgcgagaaac gtattccaac ctgcgttgcc gtttgcgtcc cctctcctgg
     5341 cactgtgcat actgccttgt tacaccctac aaccctttct caatcacgct cttcatcaga
     5401 agcaaaaatg cttattatcc acaccgcatg aaccggttct agataagatt acggtacctt
     5461 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5521 gactggaagg gctaattcac tcccaaagaa gacaagatct gctttttgcc tgtactgggt
     5581 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5641 ttaagcctca ataaagcttc agctgctcga gctagcagat ctttttccct ctgccaaaaa
     5701 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     5761 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     5821 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     5881 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     5941 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     6001 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     6061 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     6121 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6181 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6241 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6301 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     6361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     6541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6661 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccg
//
