LOCUS       sequence-490722-        8040 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_067, derived from human fusion protein FUS_FLI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8040)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_067
FEATURES             Location/Qualifiers
     source          1..8040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1573..2283,2284..2286,2287..2289))
                     /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(1615..1636)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1964..1983
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2223..2244
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2286..2295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2371..2574)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2400..2420)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2575..2878
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3135..3179)
                     /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    3364..3597
                     /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    3957..4074
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4566..4691
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4738..4995
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4997..5376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5407..5511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5512..5580,5581..6372)
                     /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(5730..5749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6543..7131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7032..7051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7285..7302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7370..7504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7394..7413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7448..7467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7510..7705)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7524..7659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7764..7785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7800..7805,7806..7823,7824..7830)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7838..7854
                     /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     7843..7865
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7862..7878
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7862..7878
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca ggggttagca agtcttgaag acgtaggtcc
      601 caagatctgg tacggatcgg gttggggtct ctgttctgtg ttcttgctaa tggtttgagc
      661 gccccccaat gggggtgact tattcaagcc agagttcatg ttgttgcccc acgcacctct
      721 tctaacagag tcataagacg ggtcaccaaa tttattgaag ccccctctgt cggagccccc
      781 cattcccccg cgtccacccc ttccaccgcc tcggcctctc ggctcatacc caccactgga
      841 cctattgtat ccaccccctc cacccccgcc gccaccacca ctgccccccc tgcctctccc
      901 acccctgtcc tgctgtccat acccaccgct gcctccgccg ccagattgat cttgatttcc
      961 gtagcccccg ccggagcctc caccagaaga cattgaggac tggtcttgtc cgtagtttcc
     1021 gcccccaccg ccgccagagc tggaattata ttggttttgc tgcccgtagc cttggggagg
     1081 attataggat tgctgctgac cgtaggactg ctgttggcct ccataactgg gttgttggga
     1141 gtagctccca ctctgaggct ggccgtatga actactttgg gaactactcc cgtaactacc
     1201 acttgtggag gatggggctg gttgctgacc ataacctgga tatgaagatt gttgaccgta
     1261 ggatgattga gaagattggc ttgatccgta accgccagtg cttccgtacc cctgtggggt
     1321 tgactgagtg ccgtatcccg tattctggct ttgcccgtat gagctgtaag aactttggcc
     1381 gtatccggag gtgtcggtgg actgactata ccctgagtag gattgctgac cgtatggctg
     1441 gctagactgt tggctgtatc cctgtccggg ttgtgtgggg tatgccccat aagattgcgt
     1501 tgcttgctgg gtataatcat ttgatgccat ccgggaagcg gccgcacttc ccccggagcc
     1561 tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     1621 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt
     1681 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1741 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     1801 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     1861 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1921 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1981 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2041 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     2101 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2161 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2221 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2281 gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagaattcg
     2341 gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct cccaccgtac
     2401 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     2461 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     2521 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     2581 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     2641 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     2701 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     2761 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     2821 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     2881 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagc
     2941 acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg aaacgataat
     3001 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     3061 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     3121 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     3181 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     3241 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     3301 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     3361 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     3421 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     3481 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     3541 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     3601 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     3661 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     3721 tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc ctgtctactt
     3781 gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt tgcttttctt
     3841 cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc acctttccag
     3901 aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa
     3961 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
     4021 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
     4081 atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct tacctgtcct
     4141 ataattttct ttaattcttt attcatagat tctactactc cttgactttg gggattgtag
     4201 ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact tttgttttgc
     4261 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
     4321 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
     4381 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagta
     4441 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
     4501 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
     4561 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
     4621 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
     4681 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
     4741 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     4801 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
     4861 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
     4921 tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc cccagtcccg
     4981 cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     5041 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     5101 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     5161 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     5221 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     5281 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     5341 ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga
     5401 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5461 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5521 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5581 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5641 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5701 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5761 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5821 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5881 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5941 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6001 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6061 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6121 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6181 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6241 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6301 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6361 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6421 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6481 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6661 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6721 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6781 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6841 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6901 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6961 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7021 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7081 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7141 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7201 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7261 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7321 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcgg atccagacat
     7381 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     7441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     7501 agttagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7561 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     7621 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     7681 tatggttgct gactaattga gatgcggatc cgctggcacg acaggtttcc cgactggaaa
     7741 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7801 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7861 acaggaaaca gctatgacca tgattacgcc aagcttgggc tgcaggtcga gggatctcca
     7921 taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag
     7981 taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat
//
