LOCUS       sequence-407485-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, TANC2_CRB3 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  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 8307)
  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_TANC2_CRB3
FEATURES             Location/Qualifiers
     source          1..8307
                     /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(3068..3778,3779..3781,3782..3784))
                     /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(3110..3131)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3459..3478
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3718..3739
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3781..3790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3866..4069)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3895..3915)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4070..4373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4630..4674)
                     /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    4859..5092
                     /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    5452..5569
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6061..6186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6233..6490
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6492..6871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6902..7006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7007..7075,7076..7867)
                     /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(7225..7244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8038..8307,1..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 ccggttcaaa
     1801 taggcaaaca gccttggacc gtttccttgc tgtcttgtgg agcacgtgcc tcggctgcat
     1861 gtgaaaactg ttcctcagac gaggggcgat acgtgccctc ggtttgacgc ttttcacgta
     1921 atttacgcac caataaagct aacccaactg caagtaaaag cgccgctaac aaagaaaaga
     1981 ctacgatgat agctgtaata gcttcggggc gtaaatttcc atccgaagat gaggaggtag
     2041 atgacggtaa cactgtgctg ttctcgttag ccgaggttgt ctgtacgcgc aagtcttgct
     2101 gagtacgcgc tcctcccatt gactgacgag gcatacgaag agatgtacga agaggagtat
     2161 gacgctgctc atcaagatat gccaggtcct ccaggtgagc gctactcgta gcggcaactg
     2221 aatttggacg cggcatcaga tataatggaa tcgattgaac actctgcgac agtacggttt
     2281 ctaagttacg ctctggaatt ttgttcagac tataggtgct agctgtcata ttctcatcca
     2341 cgcggtacag gcaactgtcc atgctgctct tttggctctg ccaaggcttg atattgccgc
     2401 ccgaacccag ctccttagat gattctcctg cgtacttggt acgctcgaca ttttccgagt
     2461 acgacgttaa cgtggcgatg ataccgctgt cgcgactctc cagagtagac gtgggggatg
     2521 taaccgcacc gtagctgcaa tctttagccg tcgcgttcgt gctaatgggc ggcaaagtag
     2581 aacagggtga ggacgccggc gggctagcag tgctagaagt caatgtcgag catggggaaa
     2641 tacgctgcgt aattgcacta gtctggtttt catcctggac ttccatcgaa tattgatccc
     2701 caggttggac ttctgtgacc ttctcgccta acaggaagcc taagcgtgtc ataagagtgt
     2761 tagctgcctc atctacggaa ggcggaggcc ccaattcctg ctcggcgtca ccttccgaca
     2821 cgggcaaggg cgggacggca tagtctggct caaaagcgca gtcagactca gtggagatgc
     2881 ccccttgacc gctgcgggac tggcgcgagt ctaccgaaga ttgacgacgg tctggaggtt
     2941 cctccgaccc cccgtcggac gagcggttct tacggctact tttgcccccc gtcaacaaca
     3001 tctttaagct attacgaaac attccccggg aagcggccgc acttcccccg gagcctccgc
     3061 ttccgccctt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     3121 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca
     3181 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     3241 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     3301 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     3361 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     3421 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     3481 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     3541 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     3601 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     3661 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     3721 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     3781 ccatggtggc gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca
     3841 ctaaacgagc ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc
     3901 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     3961 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     4021 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     4081 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4141 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4201 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4261 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4321 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4381 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg
     4441 ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga
     4501 atatccctgc ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac
     4561 caagcctcct actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa
     4621 ccaattccac aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg
     4681 ctggttttgc gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct
     4741 gtcccactcc atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact
     4801 agcattccaa ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag
     4861 gagctgttga tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat
     4921 gccccagact gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt
     4981 ctgctgctgc actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc
     5041 tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac
     5101 tgctcttttt tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg
     5161 ttcaattttt actactttat atttatataa ttcacttctc caattgtccc tcatatctcc
     5221 tccttatggt gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac
     5281 acaatcatca cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg
     5341 cactactttt atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag
     5401 ctttgctggt cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg
     5461 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     5521 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
     5581 tactgccatt tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat
     5641 tttctttaat tctttattca tagattctac tactccttga ctttggggat tgtagggaat
     5701 ttccaggtct gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc
     5761 ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata
     5821 gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg
     5881 ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac
     5941 tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt
     6001 tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact
     6061 gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact
     6121 caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg
     6181 agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag
     6241 attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga
     6301 cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc
     6361 cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag
     6421 gcaaaaagca gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag
     6481 gccacgcctc ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     6541 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     6601 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     6661 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     6721 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     6781 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     6841 tactattaat aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt
     6901 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6961 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7021 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7081 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7141 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7201 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7261 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7321 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7381 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7441 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7501 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7561 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7621 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7681 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7741 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7801 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7861 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7921 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7981 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8041 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8101 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8161 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8221 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8281 cagtggcgat aagtcgtgtc ttaccgg
//
