LOCUS       sequence-407450-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-tagged fusion protein, PTK2_PSMA7 from patient-derived
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  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 8370)
  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_PTK2_PSMA7
FEATURES             Location/Qualifiers
     source          1..8370
                     /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(3131..3841,3842..3844,3845..3847))
                     /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(3173..3194)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3522..3541
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3781..3802
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3844..3853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3929..4132)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3958..3978)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4133..4436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4693..4737)
                     /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    4922..5155
                     /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    5515..5632
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6124..6249
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6296..6553
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6555..6934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6965..7069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7070..7138,7139..7930)
                     /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(7288..7307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8101..8370,1..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 ccggttcagc
     1801 tggctttctt ttgtttcttt ttttcgtttt cttctttttc cttctcgatc tccgcgacgt
     1861 acttctcgat ttcttcagga tttaagattt tcaacgattg gtcacggcgc atcacggcta
     1921 actcgatatt cttgcctccc gattgcacaa cctcaagcag agccttaatt actaatttga
     1981 tcgtcaaatc atccgtctcg attgcttcat ccgtgtagtt cttttccaag aactcgcgga
     2041 cgctcttcgc cccgcgcccg atggcgttcg ctttccaagc gtggtatgta ccggatgggt
     2101 cggtctggta caggcgaggt gtcccatcaa aatcaaaacc cacaatcagt gcagaaatgc
     2161 caaatggacg gcggccattc gattgagtat aacgctgttt caaactcgca atgtaacgcg
     2221 tgatgtattc caccgtgact ggatcttcaa ccgtcaggcg gtgcgattgg cactctacgc
     2281 gtgcacgatt aataacgatg cgagcatcgg cagtaaggcc ggcgaaggcc atgcagacgt
     2341 tgtcgtccaa ggcacaaatt ttgcggactg tacgctcgtc ttgcaacttg gcaacactct
     2401 tcttttctac cccaagcaca acaatatcac gtccgcgaac accgactttg acaccctcat
     2461 tgtaagaatc cgcaggactg gataaactcg ctaaggaacc cagatgaccg ggggctcccg
     2521 gacgtggcgg ctttttcgga ggtgccgccg gatcgggttt ccccaccggt tggtaaatat
     2581 gctgattccc aatgggtccc tgcaaggacc catcctcgcg gtcgatactc ccacggctaa
     2641 gacgcacatc tggcttcaga aaacgctctt ctttttctaa ccaacgctga tcttcctcca
     2701 tttcctgttg ctgacggatc agacgttcct ccataagatg agtcggcagt acctgaccaa
     2761 ttccgcgaag gtccagcaca gtcgagtctt caacgtttgg ctgccacatc gcgatctcct
     2821 gtgggcggtg attccatgaa tctgtttgat ctaacaaact cgcctgtcca gggtaaattg
     2881 accctgccat cgccgtaata ccgtgactgc caggataccc ggagacttgg taatggttag
     2941 tttgaaccat gtgctggggg gaggggtaaa agccctcgct actgcgcggg gatgggtagc
     3001 ccggacgtga tggctttgga ggggcttcat cagatccacc cgaatcccaa gagacagtgg
     3061 cttggcgacg gctctccata cgcattcccc gggaagcggc cgcacttccc ccggagcctc
     3121 cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact
     3181 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc
     3241 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     3301 agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct
     3361 tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact
     3421 ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc
     3481 ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt
     3541 ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt
     3601 cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg
     3661 tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca
     3721 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     3781 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     3841 tcaccatggt ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt
     3901 tcactaaacg agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     3961 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     4021 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4081 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4141 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4201 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4261 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4321 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4381 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4441 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac
     4501 gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     4561 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     4621 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     4681 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     4741 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     4801 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     4861 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     4921 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     4981 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     5041 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     5101 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     5161 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     5221 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     5281 tcctccttat ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc
     5341 cacacaatca tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct
     5401 tggcactact tttatgtcac tattatcttg tattactact gccccttcac ctttccagag
     5461 gagctttgct ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt
     5521 ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc
     5581 tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat
     5641 gaatactgcc atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat
     5701 aattttcttt aattctttat tcatagattc tactactcct tgactttggg gattgtaggg
     5761 aatttccagg tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc
     5821 ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca
     5881 atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga
     5941 tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt
     6001 aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt
     6061 ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat
     6121 actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt
     6181 actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg
     6241 tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta
     6301 gagattttcc acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac
     6361 agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg
     6421 ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta
     6481 caggcaaaaa gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc
     6541 caggccacgc ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     6601 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     6661 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     6721 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     6781 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     6841 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     6901 gattactatt aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa
     6961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8341 tgccagtggc gataagtcgt gtcttaccgg
//
