LOCUS       sequence-408163-        7794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, S100A9_KRT4 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  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 7794)
  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_S100A9_KRT4
FEATURES             Location/Qualifiers
     source          1..7794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(113..217)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        248..627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             629..886
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    933..1058
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1550..1667
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2027..2260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2445..2489
                     /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        2746..3049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3050..3253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3204..3224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3329..3338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3335..3337,3338..3340,3341..4051)
                     /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(3380..3401)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3641..3660)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3988..4009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5059..5078)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5124..5179
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5178..5197)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5540..5556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5540..5556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5553..5575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5564..5580
                     /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(5588..5594,5595..5612,5613..5618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5633..5654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5713..5908
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5759..5894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5914..6048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5951..5970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6005..6024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6116..6133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6287..6875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6367..6386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7046..7794,1..43,44..112))
                     /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     7669..7688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
       61 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      241 ctgggtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      481 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      541 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      601 tacgtattag tcatcgctat taccatggga ggcgtggcct gggcgggact ggggagtggc
      661 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
      721 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      781 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      841 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
      901 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
      961 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1021 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1081 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1141 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatactggc ctgttagaaa
     1201 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1261 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1321 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1381 aaaaagcaca gcaagcagca ggatcttcag acctggaaat tccctacaat ccccaaagtc
     1441 aaggagtagt agaatctatg aataaagaat taaagaaaat tataggacag gtaagagatc
     1501 aggctgaaca tcttaagaca gcagtacaaa tggcagtatt catccacaat tttaaaagaa
     1561 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     1621 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     1681 gggacagcag aaatccactt tggaaaggac cagcaaagct cctctggaaa ggtgaagggg
     1741 cagtagtaat acaagataat agtgacataa aagtagtgcc aagaagaaaa gcaaagatca
     1801 ttagggatta tggaaaacag atggcaggtg atgattgtgt ggcaagtaga caggatgagg
     1861 attagaacat ggaaaagttt agtaaaacac cataaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga ccgagctcaa gcttcgaacg cgtgctagtt attaatagta
     2701 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2761 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2821 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2881 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2941 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     3001 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     3061 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3121 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3181 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3241 tataagcaga gctcgtttag tgaaccgggc tcgtttagtg aaccgaattc tgcagtcgac
     3301 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     3361 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3421 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3481 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3541 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3601 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3661 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3721 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3781 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3841 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3901 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     3961 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4021 gatcactctc ggcatggacg agctgtacaa gggcggaagc ggaggctccg ggggaagtgc
     4081 ggccgcttcc cggggaatga cgtgcaaaat gtctcagtta gaacgcaata tcgaaaccat
     4141 tatcaatacc ttccatcagt actcggttaa actgggccac cccgataccc ttaaccaagg
     4201 tgaatttaaa gaattagtgc gtaaggacct tcaaaacttt ctgaaagttc aacagttgca
     4261 aattagcgtg gaccagcacg gtgacaacct taaaaatacg aagtccgaga ttgcggaact
     4321 taaccgtatg attcaacgtc ttcgtgctga gattgaaaat atcaaaaaac aatgtcaaac
     4381 tttgcaggtc agtgtagctg atgcagaaca gcgtggggag aatgccttaa aagacgccca
     4441 ctctaaacgt gtcgagttgg aagcagcctt gcagcaagcc aaggaagaac tggcgcgtat
     4501 gttgcgcgag taccaggagt tgatgtcagt caagctggcg cttgacattg agattgctac
     4561 ctaccgcaaa ttgcttgagg gtgaagagta tcgcatgtcc ggggaatgcc aaagcgcagt
     4621 gagcatttcg gtcgtcagcg ggtcaacctc cactggcggt atttctggcg gtctgggatc
     4681 gggcagtggc tttggtcttt cgtcagggtt tggttcagga agcggctcag gctttgggtt
     4741 cggggggtcc gtcagtggct ccagttcctc taaaatcatt tcgacaacta ccctgaacaa
     4801 gcgtcgctga accggttcta gataagatta cggtaccttt aagaccaatg acttacaagg
     4861 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     4921 cccaaagaag acaagatctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     4981 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttca
     5041 gctgctcgag ctagcagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     5101 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     5161 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     5221 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     5281 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     5341 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     5401 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     5461 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     5521 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5701 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5761 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5821 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5881 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     5941 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6001 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     6061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca ccca
//
