LOCUS       sequence-407448-        7779 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express mEGFP-tagged fusion protein, MYH10_CDKN2A from 
            patient-derived sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7779)
  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 7779)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_MYH10_CDKN2A
FEATURES             Location/Qualifiers
     source          1..7779
                     /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(2540..3250,3251..3253,3254..3256))
                     /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(2582..2603)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2931..2950
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3190..3211
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3253..3262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3338..3541)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3367..3387)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3542..3845
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4102..4146)
                     /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    4331..4564
                     /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    4924..5041
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5533..5658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5705..5962
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5964..6343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6374..6478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6479..6547,6548..7339)
                     /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(6697..6716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7510..7779,1..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 catgcctgga tcgacagacc cacaggtgat tcccaatcat ctcggacgga ccctcggctg
     1861 cgtcaattcg ggcatggttg gagcctcggg tgccaccagc ggcggctcgc aaatatctag
     1921 ccacatccct atggccaagt tcctcagcca gatctaccgg caatcgcccc catgcatctc
     1981 gaacgtcaag ccgagcacca gcgcggtgca atactacgag cgtgtcgagg aagccttccc
     2041 tagccgcatc atgaactggt cgcgtcaaag ttgcaggatc tgcgcagttg ggttctgcgc
     2101 cgtgcagcag aagaagttct gcaacccgtg cggaacccat catcataaca tatatcaggc
     2161 cactataata gcgatctttc aaattatgca acacagaggc ctcgttcaaa catgtaagtt
     2221 ctgccatgtc ctccaccttg gagaatttcg gggggttcat tttctgaata tcatccttat
     2281 tgaccatagc tttctttccg ttctcggcaa gttcaaccat cacttcgtca ccccgctctt
     2341 cttttatgga ggcagcctcg aagccgtgcc gttcgctcgg tatccaaaca agtttttttg
     2401 cggtccaatc cgcttgagtg gcggggttat atatgaccgc tctatccacg aagaggtagc
     2461 gctcggggtc ctcaagtccg gtcctctggg ccattccccg ggaagcggcc gcacttcccc
     2521 cggagcctcc gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     2581 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg
     2641 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     2701 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     2761 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     2821 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     2881 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     2941 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3001 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3061 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     3121 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     3181 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     3241 cgcccttgct caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca
     3301 gaattcggtt cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc
     3361 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     3421 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     3481 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     3541 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     3601 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     3661 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     3721 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3781 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3841 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     3901 aactagcacg cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa
     3961 cgataatggt gaatatccct gcctaactct attcactata gaaagtacag caaaaactat
     4021 tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt
     4081 tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc
     4141 attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt
     4201 gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt
     4261 attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc
     4321 ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg
     4381 gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct
     4441 cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag
     4501 cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc
     4561 tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc
     4621 tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc
     4681 cctcatatct cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg
     4741 tctacttgcc acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc
     4801 ttttcttctt ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc
     4861 tttccagagg agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac
     4921 ccgaaaattt tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc
     4981 tattatgtct actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa
     5041 attgtggatg aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac
     5101 ctgtcctata attttcttta attctttatt catagattct actactcctt gactttgggg
     5161 attgtaggga atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt
     5221 gttttgctct tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg
     5281 atgcacacaa tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt
     5341 ctgaagggat ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag
     5401 gccagtatta actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt
     5461 atattttttc tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc
     5521 ccgcttaata ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat
     5581 ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg
     5641 agtcctgcgt cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg
     5701 ccactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
     5761 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
     5821 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
     5881 gacccagtac aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc
     5941 agtcccgccc aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc
     6001 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     6061 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     6121 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     6181 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     6241 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     6301 cccgtaattg attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt
     6361 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6421 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6661 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6721 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6781 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7021 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7081 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7321 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7381 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7441 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7501 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7561 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7621 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7681 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7741 agtggctgct gccagtggcg ataagtcgtg tcttaccgg
//
