LOCUS       sequence-488125-        7479 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_100, derived from human fusion protein MBNL1_MEIS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7479)
  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_100
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(383..402)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    448..503
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(502..521)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(864..880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(864..880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(877..899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    888..904
                     /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)."
     promoter        complement(join(912..918,919..936,937..942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    957..978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1037..1232
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1083..1218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1238..1372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1275..1294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1329..1348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1440..1457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1611..2199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1691..1710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2370..3161,3162..3230))
                     /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     2993..3012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3231..3335)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3366..3745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3747..4004
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4051..4176
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4668..4785
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5145..5378
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5563..5607
                     /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"
     enhancer        5864..6167
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6168..6371
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6322..6342
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6447..6456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6453..6455,6456..6458,6459..7169)
                     /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(6498..6519)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6759..6778)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7106..7127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaacg cggcattttt ccgaaagtag caaccaacat tatgcgcgct tggttgttcc
       61 aacaccttac gcatccgtac ccgtctgaag agcagaaaaa acaattagca caagataccg
      121 gcctgaccat ctgaaccggt tctagataag attacggtac ctttaagacc aatgacttac
      181 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
      241 cactcccaaa gaagacaaga tctgcttttt gcctgtactg ggtctctctg gttagaccag
      301 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
      361 ttcagctgct cgagctagca gatctttttc cctctgccaa aaattatggg gacatcatga
      421 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
      481 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
      541 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
      601 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
      661 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
      721 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
      781 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
      841 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1021 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1081 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1141 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1201 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     1261 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1321 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     1381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2041 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3361 gggtcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     3421 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3481 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3541 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3601 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     3661 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     3721 cgtattagtc atcgctatta ccatgggagg cgtggcctgg gcgggactgg ggagtggcga
     3781 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     3841 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     3901 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     3961 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     4021 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     4081 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     4141 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     4201 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     4261 atagtatggg caagcaggga gctagaacga ttcgcagtta atactggcct gttagaaaca
     4321 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     4381 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     4441 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagaaa
     4501 aaagcacagc aagcagcagg atcttcagac ctggaaattc cctacaatcc ccaaagtcaa
     4561 ggagtagtag aatctatgaa taaagaatta aagaaaatta taggacaggt aagagatcag
     4621 gctgaacatc ttaagacagc agtacaaatg gcagtattca tccacaattt taaaagaaaa
     4681 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     4741 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     4801 gacagcagaa atccactttg gaaaggacca gcaaagctcc tctggaaagg tgaaggggca
     4861 gtagtaatac aagataatag tgacataaaa gtagtgccaa gaagaaaagc aaagatcatt
     4921 agggattatg gaaaacagat ggcaggtgat gattgtgtgg caagtagaca ggatgaggat
     4981 tagaacatgg aaaagtttag taaaacacca taaggaggag atatgaggga caattggaga
     5041 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     5101 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     5161 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     5221 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     5281 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     5341 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     5401 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     5461 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     5521 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     5581 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     5641 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     5701 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     5761 ctcccaaccc cgaggggacc gagctcaagc ttcgaacgcg tgctagttat taatagtaat
     5821 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5881 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     5941 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     6001 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     6061 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     6121 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     6181 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     6241 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     6301 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     6361 taagcagagc tcgtttagtg aaccgggctc gtttagtgaa ccgaattctg cagtcgacgg
     6421 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     6481 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     6541 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     6601 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     6661 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     6721 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     6781 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     6841 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     6901 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6961 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     7021 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     7081 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     7141 tcactctcgg catggacgag ctgtacaagg gcggaagcgg aggctccggg ggaagtgcgg
     7201 ccgcttcccg ggttattgat gaacgcgatg gcagtagcaa atctgatcac gaggaacttt
     7261 cggggtcgag cacgaacctt gccgaccata atcctagttc ctggcgcgat cacgatgacg
     7321 ctacctcgac acacagcgcg ggaacacccg gtccttcttc ggggggtcat gcatcccagt
     7381 cgggagacaa ctcatctgaa cagggagatg ggcttgacaa ttccgtggca tcaccgggca
     7441 ccggtgatga tgacgaccca gataaggata aaaaacgtc
//
