LOCUS       sequence-407436-        8496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PCBD2_KRT13 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8496)
  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 8496)
  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_PCBD2_KRT13
FEATURES             Location/Qualifiers
     source          1..8496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    558..692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(582..601)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     636..655
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(698..893)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      712..847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    952..973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(988..993,994..1011,1012..1018)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1026..1042
                     /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     1031..1053
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1050..1066
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1050..1066
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1409..1428
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1427..1482
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1528..1547
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3257..3967,3968..3970,3971..3973))
                     /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(3299..3320)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3648..3667
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3907..3928
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3970..3979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4055..4258)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4084..4104)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4259..4562
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4819..4863)
                     /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"
     misc_feature    5048..5281
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5641..5758
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6250..6375
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6422..6679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6681..7060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7091..7195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7196..7264,7265..8056)
                     /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(7414..7433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8227..8496,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
      601 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      661 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
      721 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
      781 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
      841 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg
      901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      961 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1021 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1081 gcttgggctg caggtcgagg gatctccata agagaagagg gacagctatg actgggagta
     1141 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     1201 aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc
     1261 tatggaataa ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc
     1321 aacctttgtt catggcagcc agcatatggg catatgttgc caaactctaa accaaatact
     1381 cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa
     1441 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
     1501 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct gctagctcga
     1561 gcagctgaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     1621 ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagat cttgtcttct
     1681 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     1741 agctgccttg taagtcattg gtcttaaagg taccgtaatc ttatctagaa ccggttcatg
     1801 gacgacggac atccgacgta cggcgtccgc ttgcgttact ggtcgtggtg actgacgcac
     1861 tggatgtggt cgtgactgat gtagagcgtg gagacacact accggcggag gaaggaaatc
     1921 cgatcatctt agcatcctgc ccttccaaca atgaacgata cgtagcaatc tcttgttcta
     1981 aacgtgtttt aatgtcaagt aacatcttgt actcttggtt ttggcactcc atctcggagc
     2041 gaagctcact taattgggct tcaatggagg agatcaaacc ctgaatttgt tgaagctgca
     2101 gcgcgtagcg gcattccgtt tctgcaactg tgttttccag ccctgctttc atgctcagct
     2161 ggctttgtaa ttcaatctct aaaccctgta aagtgcggcg aagctccgta atttccgttt
     2221 tagacgtttg aatcatagcg gtattagttg aaacttcctt attcagttcc gctgacttag
     2281 tatggaacca ttcctcagca tcgcggcggt tgcgttcggc catagcttcg tattgttcac
     2341 gcatctcggc caggacgcgg gttaaatcaa tccctggagt agcatccatc tcaacattaa
     2401 cctgaccgac cacctgatta gaaaattctt tcatttcctc ttcatgattc ttcttcatgt
     2461 aggccaactc ctcgttaagc gattcaatct gcatctccaa atcagtctta ctcagagtaa
     2521 gctcgtccag tacgcggcgc aacccattga tgtcggcttc cacagactgg cgcagcgcca
     2581 gctcattctc atattttaag cggaagtcat cggcggcaag acgtgcgttg tcgatttcca
     2641 aaatcacacg attgttttca atagtagcag taaggatttt gtcacgcaac tcttcgatcg
     2701 tcttgtagta tggactataa tcgcgctctg gactagccgg actttgcttt aaatgccagt
     2761 cacggatttt tacctcaaga tcggcgtttg cctcttccaa ggcgcgcacc ttctctaagt
     2821 agctagccaa acgatcatta agattctgca tcgtgatttt ttcattccct gtaagcaacc
     2881 ctccatcgca ggccccgaag tctacaaagc cacctgcgaa cccaccgccg aagcctcctc
     2941 caaggccccc accataccca ccacccagtc cgccgccgta accaccacca aaaccctgat
     3001 taaaattatg gaacgaaaat tccttgtaga tggcatcacg ttccgacagc tctgaccagc
     3061 cagccgcctt caagtccaag atggcttggt tacgttcttc ggcagtcagg cggtgtgtcc
     3121 cggacgacat agccgccagt cccaaagact gtccgcgtaa ggcggcaagc aagcgacggg
     3181 tggcaccaag ggctcccaac actgctgcca ttccccggga agcggccgca cttcccccgg
     3241 agcctccgct tccgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3301 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     3361 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3421 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     3481 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     3541 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3601 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     3661 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     3721 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     3781 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     3841 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     3901 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     3961 ccttgctcac catggtggcg accggtggat cccgggcccg cggtaccgtc gactgcagaa
     4021 ttcggttcac taaacgagcc cggttcacta aacgagctct gcttatatag acctcccacc
     4081 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     4141 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     4201 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     4261 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     4321 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     4381 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     4441 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     4501 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     4561 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     4621 tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt gggaggtggg tctgaaacga
     4681 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     4741 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     4801 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     4861 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     4921 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     4981 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     5041 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     5101 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     5161 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     5221 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     5281 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     5341 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     5401 catatctcct ccttatggtg ttttactaaa cttttccatg ttctaatcct catcctgtct
     5461 acttgccaca caatcatcac ctgccatctg ttttccataa tccctaatga tctttgcttt
     5521 tcttcttggc actactttta tgtcactatt atcttgtatt actactgccc cttcaccttt
     5581 ccagaggagc tttgctggtc ctttccaaag tggatttctg ctgtccctgt aataaacccg
     5641 aaaattttga atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat
     5701 tatgtctact attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt
     5761 gtggatgaat actgccattt gtactgctgt cttaagatgt tcagcctgat ctcttacctg
     5821 tcctataatt ttctttaatt ctttattcat agattctact actccttgac tttggggatt
     5881 gtagggaatt tccaggtctg aagatcctgc tgcttgctgt gcttttttct tacttttgtt
     5941 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
     6001 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
     6061 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
     6121 agtattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
     6181 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
     6241 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
     6301 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
     6361 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
     6421 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
     6481 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
     6541 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
     6601 ccagtacagg caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt
     6661 cccgcccagg ccacgcctcc catggtaata gcgatgacta atacgtagat gtactgccaa
     6721 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     6781 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     6841 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     6901 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     6961 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     7021 gtaattgatt actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg
     7081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     7141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     7201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     7261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     7321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     7381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     7501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     7561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     7621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     8041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     8101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8461 ggctgctgcc agtggcgata agtcgtgtct taccgg
//
