LOCUS       sequence-406620-        7848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, CREBBP_ZNF500 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  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 7848)
  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_CREBBP_ZNF500
FEATURES             Location/Qualifiers
     source          1..7848
                     /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(2609..3319,3320..3322,3323..3325))
                     /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(2651..2672)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3000..3019
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3259..3280
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3322..3331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3407..3610)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3436..3456)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3611..3914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4171..4215)
                     /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    4400..4633
                     /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    4993..5110
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5602..5727
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5774..6031
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6033..6412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6443..6547
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6548..6616,6617..7408)
                     /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(6766..6785)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7579..7848,1..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 ctttggctcc tggcccgccc ggagccaccg gttgcagagt cggaagtgaa cctgccccca
     1861 tgtgggtgcg ttgatgcttg tgaagatctg ttccacggcg gaacccacgt ccacaagcgg
     1921 gacatgtgta gggcttttcc cccgtatgtg tacgacggtg agcactgaaa tgtgatgaat
     1981 tattaaaacg cttcccgcat tgagtacagg catacggacg ctctccagaa tgggtgcgac
     2041 ggtgaattac aagagaactg gactgactaa agcgtttccc acactctggg cagggatagg
     2101 gcttctcccc cgtgtggacg cgttggtggg ttgaaaaatt cgagcgatca ctaaatcctt
     2161 taccgcacac caaacatttg tagggacgct ccccagtgtg agtacgctgg tgtttggtta
     2221 agtgcgaagt cttgctgaag cccttaccgc attcagggca agtatagggc ttgtcagccc
     2281 cgtggctcgc gcgacgaccc ggtgggggcc ctccacgtgg ttggtctggg cgtactaaac
     2341 cacgtacggg agccggacgt tgcccaggaa ggggatgacc gggaccctga caacgtgctg
     2401 ataacacacg ataccattcc atgcgtaatg gtgcgtcttc gcgtccatca ccgccatcct
     2461 caagttggat tcccggaccc gtactatcgt tggcggagaa acctggagag ctcagcttgg
     2521 cgcgcttagg attaggcggt ccgtctaaca agttttcagc cattccccgg gaagcggccg
     2581 cacttccccc ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc
     2641 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2701 tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2761 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     2821 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     2881 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     2941 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3001 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3061 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     3121 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     3181 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3241 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3301 acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg
     3361 tcgactgcag aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat
     3421 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     3481 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     3541 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     3601 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3661 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3721 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3781 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     3841 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     3901 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     3961 actattaata actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg
     4021 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
     4081 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
     4141 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
     4201 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
     4261 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
     4321 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
     4381 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
     4441 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
     4501 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     4561 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     4621 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     4681 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     4741 ccaattgtcc ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc
     4801 ctcatcctgt ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat
     4861 gatctttgct tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc
     4921 cccttcacct ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct
     4981 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
     5041 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
     5101 tcttttaaaa ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg
     5161 atctcttacc tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg
     5221 actttgggga ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt
     5281 cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc
     5341 tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc
     5401 tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt
     5461 ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg
     5521 ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct
     5581 aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct
     5641 agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc
     5701 agcaagccga gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct
     5761 gttcgggcgc cactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     5821 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     5881 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     5941 aaccagagag acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg
     6001 ccactcccca gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag
     6061 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6121 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     6181 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     6241 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     6301 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     6361 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg
     6421 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6481 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6541 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6601 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6661 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6721 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6781 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6841 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6901 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6961 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7021 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7081 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7141 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7201 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7261 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7321 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7381 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7441 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7501 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7561 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7621 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7681 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7741 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7801 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgg
//
