LOCUS       sequence-406466-        8679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, AIMP2_EIF2AK1 from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8679)
  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 8679)
  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_AIMP2_EIF2AK1
FEATURES             Location/Qualifiers
     source          1..8679
                     /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(3440..4150,4151..4153,4154..4156))
                     /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(3482..3503)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3831..3850
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4090..4111
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4153..4162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4238..4441)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4267..4287)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4442..4745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5002..5046)
                     /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    5231..5464
                     /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    5824..5941
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6433..6558
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6605..6862
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6864..7243
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7274..7378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7379..7447,7448..8239)
                     /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(7597..7616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8410..8679,1..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 ttatctagat caacccaccc
     1801 cgccgtcttt tccatcatca cgcacgccct tgtcctgcga caaaagattc aattgtttct
     1861 taagctcagc gatctctttt tcctgctcaa taatcttcat ttgcaacgtc aggtttacat
     1921 taccgctatt ttgaaacaac tcagactgta ataactgaat cgcactgggg cgctgagacg
     1981 agttgcggcg tgtcagatgc tggatatatt tagcctgcac aggacaacgc ttgcgaagcg
     2041 actccggcag ttggccggta cgcagtcctg taagcacttc cgcgcgttcc atttcagtgc
     2101 cgaaaggttg gaacagctcc aacagcacaa cccccagcga gtacatatct gacttggcgt
     2161 catactccga gccctccagt tgctctgggc tagcatacag gcaggtgcca acacgtgaag
     2221 tatgtgtggg tgtacgcttg ccattgcgat tcgtccagtc agtgttcttc tgcagaatgt
     2281 cggtacaagc caggccgaag tcaccaattt ttacttgctg atcagggccg tgtaaaaaaa
     2341 tattacgtgg tttcaggtcg cggtgaacaa tacccatatt gtggatgtag aagacacctt
     2401 ccaccaattc ttgaaagatt ttggtggcaa cgtttgccat tacatagggg catgcggact
     2461 cgtctacata ctcgcgcccg cgtttgttac gttccacaat ccagtcccac aagctcagtt
     2521 cacacagttg catctggatg tgaagcataa ggtgatattg tgcctcggtc tgccccaaaa
     2581 aatttacgtt ttcctcactc gattcctcgg tagatgtgaa agattcctca agatgagaat
     2641 tacgacgtaa tggaagctgt tgctcgacga tagacgaagc gctaagtcca gctaacccat
     2701 tttcttgaag ttccaacgtt gactccagtt ctccgctttc acgaataact aagttcgtag
     2761 tgtactttac tgatttgttg ttctggtttt ctgtatcact ctctccaaag cgcttctctt
     2821 tctcaggcgt aggttccgcg aaaataatcg aggaagagga tgactcgtca ttcttcacgc
     2881 cacactgctc gcggtcctct tcttggtcac ttaaaacctc taacgaaggc aactcaattg
     2941 ctgcgcggtc ggcacgaggc tggattacat ggacgtgttc gatccacgca gtatggtaac
     3001 cgacaatatt agggtgttgc agcccggcca aaactttgac ttcacgcaag acctttccta
     3061 acacggaatt taagtccaga gcattggtcg taagggtggt gggctcatca gcctgaataa
     3121 tattggtgac gtccaagtca gcatctggtg tttggatcat cttcgacaat ccatcgactg
     3181 cagccttcag ctcatacaga cgtttcaaaa tgtcatcctg acgtgattct aatgcttgca
     3241 aagaaagatt cgattcttcc tgcacatgac ctgcacctgg cgccggacca tagctgcgac
     3301 catgaacgtt aggtaaacgg tacatacaag tcggcagctc cacacgcagc ggtgcccctc
     3361 ctccatgata tggctttacc tgatacatag gcattccccg ggaagcggcc gcacttcccc
     3421 cggagcctcc gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     3481 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
     3541 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     3601 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     3661 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     3721 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3781 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3841 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3901 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3961 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     4021 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     4081 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     4141 cgcccttgct caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca
     4201 gaattcggtt cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc
     4261 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     4321 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     4381 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     4441 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     4501 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     4561 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     4621 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     4681 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     4741 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     4801 aactagcacg cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa
     4861 cgataatggt gaatatccct gcctaactct attcactata gaaagtacag caaaaactat
     4921 tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt
     4981 tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc
     5041 attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt
     5101 gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt
     5161 attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc
     5221 ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg
     5281 gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct
     5341 cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag
     5401 cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc
     5461 tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc
     5521 tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc
     5581 cctcatatct cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg
     5641 tctacttgcc acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc
     5701 ttttcttctt ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc
     5761 tttccagagg agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac
     5821 ccgaaaattt tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc
     5881 tattatgtct actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa
     5941 attgtggatg aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac
     6001 ctgtcctata attttcttta attctttatt catagattct actactcctt gactttgggg
     6061 attgtaggga atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt
     6121 gttttgctct tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg
     6181 atgcacacaa tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt
     6241 ctgaagggat ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag
     6301 gccagtatta actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt
     6361 atattttttc tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc
     6421 ccgcttaata ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat
     6481 ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg
     6541 agtcctgcgt cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg
     6601 ccactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
     6661 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
     6721 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
     6781 gacccagtac aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc
     6841 agtcccgccc aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc
     6901 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     6961 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     7021 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     7081 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     7141 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     7201 cccgtaattg attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt
     7261 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7321 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7381 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7441 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7501 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7561 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7621 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7681 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7741 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7801 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7861 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7921 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7981 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     8041 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     8101 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8161 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8221 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8281 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8341 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8521 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8641 agtggctgct gccagtggcg ataagtcgtg tcttaccgg
//
