LOCUS       sequence-407520-        8076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, KLK3_KLK2 from patient-derived 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8076)
  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 8076)
  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_KLK3_KLK2
FEATURES             Location/Qualifiers
     source          1..8076
                     /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(5341..5360)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5406..5461
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5460..5479)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5822..5838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5822..5838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5835..5857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5846..5862
                     /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(5870..5876,5877..5894,5895..5900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5915..5936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5995..6190
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6041..6176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6196..6330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6233..6252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6287..6306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6398..6415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6569..7157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6649..6668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7328..8076,1..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     7951..7970
                     /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 cggggaatgt gggtgccagt ggtgtttctg accctttctg taacgtggat
     4141 cggggccgct ccccttatcc tgtcgcgcat cgtgggcggc tgggaatgtg aaaaacattc
     4201 acaaccctgg caggtacttg tcgcatcccg tgggcgtgct gtctgtgggg gtgtcttagt
     4261 ccacccacag tgggtgctta ccgcagctca ttgtatccgt aataagtccg taaacctggt
     4321 gttaagtatt gcgctttcgg ttggatgcac gggtgcagta ccacttattc aatcccgcat
     4381 cgtaggcggg tgggaatgcg agaagcacag tcagccctgg caggttgcgg tgtattctca
     4441 cggatgggcc cattgtggcg gagttctggt tcatccccaa tgggtgttaa cagcagctca
     4501 ttgcttaaag aaaaactcac aggtctggtt gggtcgtcat aatctgttcg agcctgagga
     4561 tacaggacaa cgcgtcccgg taagccattc ctttcctcat cctttgtata acatgtcgtt
     4621 acttaagcat cagtcgttac gtcctgatga agacagtagc cacgacttga tgttgcttcg
     4681 cttgagcgaa cctgcaaaga tcacagacgt agtgaaggtg ctgggcctgc ctacgcagga
     4741 acctgccctt gggaccactt gctatgcttc cggatggggg tccatcgagc ctgaggagtt
     4801 tcttcgccct cgctcgcttc agtgtgtctc attacattta ctttccaacg atatgtgcgc
     4861 tcgcgcgtat agcgaaaaag taaccgaatt tatgttgtgc gcaggtctgt ggacaggagg
     4921 taaagacaca tgcggaggcg atagcggcgg gccacttgta tgcaatggag tattacaagg
     4981 gattaccagt tgggggcccg aaccatgtgc actgcctgag aagcctgctg tttacacgaa
     5041 agtagtccat tatcgcaagt ggattaagga caccatcgcg gcaaacccgt gaaccggttc
     5101 tagataagat tacggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5161 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaaaga agacaagatc
     5221 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5281 gctaactagg gaacccactg cttaagcctc aataaagctt cagctgctcg agctagcaga
     5341 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     5401 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     5461 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     5521 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     5581 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     5641 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     5701 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     5761 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     5821 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5881 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5941 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     6001 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6061 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6121 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6181 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6241 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6301 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     6361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7321 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7381 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7441 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7501 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7561 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7621 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7681 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7741 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7801 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7861 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7921 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7981 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8041 tgagatccag ttcgatgtaa cccactcgtg caccca
//
