LOCUS       sequence-488144-        8994 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_121, derived from human fusion protein NUP98_KDM5A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8994)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8994)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_121
FEATURES             Location/Qualifiers
     source          1..8994
                     /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(2527..3237,3238..3240,3241..3243))
                     /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(2569..2590)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2918..2937
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3177..3198
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3240..3249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3325..3528)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3354..3374)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3529..3832
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4089..4133)
                     /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    4318..4551
                     /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    4911..5028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5520..5645
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5692..5949
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5951..6330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6361..6465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6466..6534,6535..7326)
                     /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(6684..6703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7497..8085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7986..8005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8239..8256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8324..8458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8348..8367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8402..8421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8464..8659)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8478..8613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8718..8739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8754..8759,8760..8777,8778..8784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8792..8808
                     /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     8797..8819
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8816..8832
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8816..8832
                     /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 cacagcattc tcatcatcag actcctctgc
      601 tcctgaccag tcatactttg aggggatgtc cagcaccttt ttctctcttt tcttttcagt
      661 gctctctttc acaagttcaa ctttggctgc agcagccttc tccttctttt tctttctctc
      721 ttcttctttt gctagtttct tggccagttt attcagctcc tttgatttgt ctgcacctaa
      781 ttttaatttc ttctttctag gtttgtccat tttctttaac tccttggact tctgttttcc
      841 ttctccaaaa agttgctcta ccttttctag cttccgtttc cgtttcttct ctgaagagtc
      901 ctttcctttc acttttagtg gtttctcttc catgctgtca tcctcttcct tcttcttagg
      961 gtctgacatc ggattccgga agagaggaga gtctccaaaa ggtgagtatg ttagactatt
     1021 gatgtgctgc tggagaacag cctgctgggc agcagaagca tttggatctg tcaaagctac
     1081 tggggcctgg ggggctccaa agcccaaagt ggctgtcgta gtattaaatc caggggcccc
     1141 aaaggctcct gtaccaagag gccctccaat cttaggttgg ttgttcccaa acaaagatgc
     1201 ctgtccagca ccaagagctg ttccaaatcc tgcaccaagc ccagttccaa gagtcccagg
     1261 tgctggttta cttccaaaaa tactattccc actggtattt gtgccaaaac caaagagccc
     1321 gccactggtt gtaccaaatg aaggtgcact ggttgtgctg cttccaaatg tagtaagctt
     1381 gttattgcca aacagggtcg aaccaacagc accaaatcca gtattggtct gcccaaagag
     1441 acctgttcct gttccaaatg ctgtcccagt gctggtgttt gtagctgtcc caaaaagacc
     1501 tccaggctgt gaggcttggg ttactccaaa taatcccatg gtgttggtgc ttggctgtcc
     1561 tatggtgctg gtattaccaa aggaaaagcc agtgttctgg gtggttgtag cctggccaaa
     1621 tggtttgctg aagaggctgg tagtctgctg attctgttgg ccaaagagac cacctggatt
     1681 tgttccaaat ccagttgtac tagttccaaa ggcagttttg ttctgaccat atgcaaagcc
     1741 tgaattagtg gtggaggagc tgaagagtcc tgttgcgctg gaagtggctg gagaagaccc
     1801 aaacaagcca gttgtggtac ctgctcccac ctggttctgt gggcccttcc tgttagcctg
     1861 ataatcctct aaacgaagtt cctctagtga cttgctttca tattctttca tagcagtaat
     1921 acactggtgc ttggtactta tgttagtgct aactccagct ttgaccatag tatctgtacc
     1981 agttggaggg ttaaatttaa tagtagtccc agtaggagca gctgtaaaac tacttggccc
     2041 aaagagggag ccagatgtgc tgccaaaagg attagaggtg gtatttgtgg ttccaaagag
     2101 tcctccactg ctagtactgg ttccaaaatt gccaaagcca gttggtttat tttgtgcaaa
     2161 ggcattgttt tgggatgaga agagactggt ccctgtgctt gcagttccaa acaaggtatt
     2221 tgctgttcct gttgacgtac caaacccaaa gccagtgctt gtggaggtag ctggctggct
     2281 aaatgaactg gttccaaaca atcctcctgg tttagtctgt gaatttccaa agaggcctcc
     2341 agtattgttg ctagaaccaa atgcagatgt tccaaatgcc cctccactag tagtgccaaa
     2401 gccagtattc tgtccaaatg ttgaagttgt gccaaagcca cctgtgccac ccccaaaggg
     2461 tgttccaaat gatttgttaa acatccggga agcggccgca cttcccccgg agcctccgct
     2521 tccgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     2581 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact gggtgctcag
     2641 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     2701 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     2761 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     2821 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     2881 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     2941 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     3001 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     3061 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     3121 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     3181 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     3241 catggtggcg accggtggat cccgggcccg cggtaccgtc gactgcagaa ttcggttcac
     3301 taaacgagcc cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct
     3361 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     3421 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     3481 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     3541 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     3601 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     3661 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     3721 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     3781 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     3841 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagcacgcgt
     3901 tcgaagcttg agctcggtcc cctcggggtt gggaggtggg tctgaaacga taatggtgaa
     3961 tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct taaacctacc
     4021 aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt tatgttaaac
     4081 caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt cttttcttgc
     4141 tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt aatttctctg
     4201 tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt actccaacta
     4261 gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca aatccccagg
     4321 agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag ctgcttgatg
     4381 ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag caaattgttc
     4441 tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt cattgacgct
     4501 gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc tattcccact
     4561 gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac tcctaatggt
     4621 tcaattttta ctactttata tttatataat tcacttctcc aattgtccct catatctcct
     4681 ccttatggtg ttttactaaa cttttccatg ttctaatcct catcctgtct acttgccaca
     4741 caatcatcac ctgccatctg ttttccataa tccctaatga tctttgcttt tcttcttggc
     4801 actactttta tgtcactatt atcttgtatt actactgccc cttcaccttt ccagaggagc
     4861 tttgctggtc ctttccaaag tggatttctg ctgtccctgt aataaacccg aaaattttga
     4921 atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat tatgtctact
     4981 attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt gtggatgaat
     5041 actgccattt gtactgctgt cttaagatgt tcagcctgat ctcttacctg tcctataatt
     5101 ttctttaatt ctttattcat agattctact actccttgac tttggggatt gtagggaatt
     5161 tccaggtctg aagatcctgc tgcttgctgt gcttttttct tacttttgtt ttgctcttcc
     5221 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
     5281 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     5341 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc agtattaact
     5401 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     5461 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     5521 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     5581 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     5641 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     5701 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     5761 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     5821 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg
     5881 caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt cccgcccagg
     5941 ccacgcctcc catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     6001 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     6061 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     6121 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     6181 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     6241 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     6301 actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg
     6361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8281 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcggatccag acatgataag
     8341 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     8401 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttag
     8461 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     8521 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     8581 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     8641 tgctgactaa ttgagatgcg gatccgctgg cacgacaggt ttcccgactg gaaagcgggc
     8701 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8761 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8821 aacagctatg accatgatta cgccaagctt gggctgcagg tcgagggatc tccataagag
     8881 aagagggaca gctatgactg ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac
     8941 atgtaaggaa aattttaggg atgttaaaga aaaaaataac acaaaacaaa atat
//
