LOCUS       sequence-487989-        7695 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_081, derived from human fusion protein KMT2A_AFDN; 
            KMT2A_EPS15; KMT2A_MLLT1; KMT2A_MLLT10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7695)
  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_081
FEATURES             Location/Qualifiers
     source          1..7695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(480..499)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    545..600
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(599..618)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(961..977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(961..977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(974..996)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    985..1001
                     /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(1009..1015,1016..1033,1034..1039))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1054..1075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1134..1329
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1180..1315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1335..1469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1372..1391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1426..1445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1537..1554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1708..2296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1788..1807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2467..3258,3259..3327))
                     /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     3090..3109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3328..3432)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3463..3842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3844..4101
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4148..4273
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4765..4882
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5242..5475
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5660..5704
                     /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        5961..6264
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6265..6468
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6419..6439
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6544..6553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6550..6552,6553..6555,6556..7266)
                     /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(6595..6616)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6856..6875)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7203..7224
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaaac agccgccgcc accggaaagc ggtccggagc agtcgaagca gaaaaaggtg
       61 gcaccgcgtc cgtctattcc ggtgaagcaa aaacccaaag agaaggaaaa gcctccgccc
      121 gtaaacaaac aagaaaacgc aggcaccttg aatattttgt ccaccctgag taacggtaac
      181 agcagcaagc aaaagattcc ggcggatggt gtgcaccgta tccgtgtgtg aaccggttct
      241 agataagatt acggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
      301 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaaagaa gacaagatct
      361 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      421 ctaactaggg aacccactgc ttaagcctca ataaagcttc agctgctcga gctagcagat
      481 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
      541 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
      601 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
      661 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
      721 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
      781 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
      841 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
      901 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
      961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     1141 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     1201 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     1261 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     1321 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1381 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1441 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     1501 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1561 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2461 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2521 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2581 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2641 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2701 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2761 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     2821 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2881 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2941 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3001 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3061 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3121 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3181 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3241 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3301 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3361 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3421 aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta
     3481 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3541 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3601 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3661 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3721 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3781 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3841 tgggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc
     3901 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     3961 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4021 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4081 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     4141 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     4201 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     4261 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     4321 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     4381 agaacgattc gcagttaata ctggcctgtt agaaacatca gaaggctgta gacaaatact
     4441 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     4501 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     4561 agacaagata gaggaagagc aaaacaaaag taagaaaaaa gcacagcaag cagcaggatc
     4621 ttcagacctg gaaattccct acaatcccca aagtcaagga gtagtagaat ctatgaataa
     4681 agaattaaag aaaattatag gacaggtaag agatcaggct gaacatctta agacagcagt
     4741 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     4801 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     4861 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagaaatc cactttggaa
     4921 aggaccagca aagctcctct ggaaaggtga aggggcagta gtaatacaag ataatagtga
     4981 cataaaagta gtgccaagaa gaaaagcaaa gatcattagg gattatggaa aacagatggc
     5041 aggtgatgat tgtgtggcaa gtagacagga tgaggattag aacatggaaa agtttagtaa
     5101 aacaccataa ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     5161 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     5221 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     5281 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     5341 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     5401 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     5461 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     5521 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     5581 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     5641 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     5701 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     5761 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     5821 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggaccgag
     5881 ctcaagcttc gaacgcgtgc tagttattaa tagtaatcaa ttacggggtc attagttcat
     5941 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     6001 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     6061 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     6121 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     6181 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     6241 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     6301 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     6361 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     6421 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     6481 cgggctcgtt tagtgaaccg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     6541 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6601 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     6661 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     6721 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     6781 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     6841 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     6901 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     6961 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     7021 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     7081 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7141 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     7201 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7261 tacaagggcg gaagcggagg ctccggggga agtgcggccg cttcccggtg gatgccatcc
     7321 aaggcgtacc tgcagaaaca ggccaaggca gttaagaaaa aagagaaaaa atccaagact
     7381 tctgagaaaa aggacagcaa ggagtcgagc gtcgtcaaga atgtggtaga tagctcccag
     7441 aagcccacgc cgagcgctcg tgaagacccg gcaccaaaaa agagcagctc tgagcctccg
     7501 ccgcggaaac cggtggaaga gaaaagtgaa gaaggtaatg ttagcgcacc gggcccggag
     7561 tccaagcagg ccacgactcc agctagtcgt aaatcttcca aacaggtttc acaaccggcc
     7621 ttggtcattc cgccgcagcc gccgaccaca ggtccgccgc gcaaagaggt gccgaagacc
     7681 acccctagcg agccg
//
