LOCUS       sequence-407464-        8958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, SLC16A14_SP110 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8958)
  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 8958)
  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_SLC16A14_SP110
FEATURES             Location/Qualifiers
     source          1..8958
                     /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(3719..4429,4430..4432,4433..4435))
                     /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(3761..3782)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4110..4129
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4369..4390
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4432..4441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4517..4720)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4546..4566)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4721..5024
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5281..5325)
                     /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    5510..5743
                     /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    6103..6220
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6712..6837
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6884..7141
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7143..7522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7553..7657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7658..7726,7727..8518)
                     /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(7876..7895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8689..8958,1..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 ccggttcacg
     1801 gcagagtcca gaaaccaccg tcgttggctt cgtgaaatcc aagtacgtcc ttaagatcct
     1861 tctcgaactc ggcctcaaga tccaagccga cttgcccaaa atcactagct ttatagaacg
     1921 tcttatgatt gcgaaacatt aaacgcatat cacgaacaaa ccacgcgacc gtgtacatct
     1981 cagtaatcaa acgttctttt accagatcca gccacatggc ctcctgaaat ggttccccgt
     2041 agtcgcgaat attaaacggg atgccggtga agaagctgga ctgggggtgg caataggctt
     2101 tcaacagcaa aaactcgcac ttcagttgat cttgaggttg catttggcgt tcaagggtct
     2161 tcgagacatg atgacactgc tgggaaccac tactgcgctt catgcgacag aacgtacaac
     2221 tccacagcat acgttttgct tccacgggag gaatgtgaca atcctcgtga aaaacacgag
     2281 gacacgtgcc gcaacaaaga agttgtcccc cttggcaaca gacctcacat tcatctgagt
     2341 tcttgcgttt aagcaattcg cccaacgtca tcccttcaca acgaatattg cgtttccaat
     2401 ttttcgcgtt gcggccctta ccctcgactt caaactcatt aggggtcagc caggtgccgt
     2461 cctcattgcg gatgcattta actgacgaac cgtgtttcat tttcttctta tacagaatcc
     2521 cttttgcctc tccacaggtg accggcagtt tggagcagtg aaagtctact gtatctgatt
     2581 tgggcttacc gcgacggtgg atgttcttct gaaaacggcg cttggatgag gaacagatgt
     2641 ctttttcttt cttcttctct tttaaacgtg acttgcgagc gcacttggtg cgggcttttt
     2701 ggacacgcat catcacctca ctattccacg tcgagtcgtc tttacgctgt gtgaccttgt
     2761 caacgacctg aagtttttcc tggattccat gccccggtga ggcagcccct tgggtcacgc
     2821 gacgtggtgt ggacggggtc ttctggctgc gcttgccctc attcatttcg gatgtaccat
     2881 cgataatttc ttccgaacga ctcttgcgtg cgcactcagt gcgagctttc tgcgcacgag
     2941 tctccacggt ggaattgcaa gtgctgtcat ccttcttttg ggggacctgg tctacgcgct
     3001 tcaacttttt ctggatacca tgacgggaag atgcagtccc gcccggtaaa cttttttttt
     3061 tatggcggcg cttgggcgtg ctccagatac agcgcttacg cttcttcccc ttcttatcgc
     3121 ttggggtcga gcttacctcc tggggctctt ccgggtcatt gggttccgga ctattgtcgc
     3181 ggatctcagg catagaacct aagggggaat gaggcatctc ttgagggtct tccttgtcac
     3241 gaatttgcgg gataagattg tccgaggcga cagccgccac cccgaaggta atgaaaagat
     3301 aatgaacgtt agcagcgtag gcactcagta cccatccaag agaattaaca agtcctccga
     3361 tgatggccgt ctggcggcaa ccgcacgtat tgataaataa accaataaac gggccaacaa
     3421 ttaaagtgat ccccatggac aatgacgaaa cccacgcagt cagcccgcga gattggtgaa
     3481 attcctctaa ccactcgaca tttaacacgc caagtgccat ctgggaaccc ataataagga
     3541 tgtgaacaaa gaatgaggaa agtaccatca tccaagccca ccctccatca atattcgggt
     3601 ggggtttaag cgttttcttg tctttcggtc cgtcttcaaa gtcatatcca atatcttcat
     3661 gagaagtgta cattccccgg gaagcggccg cacttccccc ggagcctccg cttccgccct
     3721 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3781 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt
     3841 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     3901 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     3961 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     4021 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     4081 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     4141 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     4201 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     4261 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     4321 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     4381 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     4441 cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag
     4501 cccggttcac taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     4561 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     4621 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     4681 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     4741 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     4801 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     4861 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     4921 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     4981 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     5041 ggctatgaac taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct
     5101 tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     5161 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     5221 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     5281 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     5341 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     5401 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     5461 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     5521 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     5581 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     5641 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     5701 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     5761 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     5821 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg
     5881 tgttttacta aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc
     5941 acctgccatc tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt
     6001 tatgtcacta ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg
     6061 tcctttccaa agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     6121 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     6181 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     6241 ttgtactgct gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa
     6301 ttctttattc atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc
     6361 tgaagatcct gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt
     6421 gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc
     6481 tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg
     6541 tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg
     6601 ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg
     6661 ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc
     6721 gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg
     6781 ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc
     6841 ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac
     6901 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     6961 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     7021 ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc
     7081 agctgcttat atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct
     7141 cccatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     7201 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     7261 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     7321 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     7381 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     7441 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     7501 taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac
     7561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     7621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     7681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     7741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     8041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     8101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     8161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     8221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     8281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     8341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     8401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     8461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     8521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     8581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     8641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     8701 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     8761 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8821 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8881 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8941 taagtcgtgt cttaccgg
//
