LOCUS       sequence-487988-        7686 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_079, 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 7686)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7686)
  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_079
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(422..441)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    487..542
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(541..560)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(903..919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(903..919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(916..938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    927..943
                     /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(951..957,958..975,976..981))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    996..1017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1076..1271
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1122..1257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1277..1411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1314..1333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1368..1387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1479..1496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1650..2238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1730..1749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2409..3200,3201..3269))
                     /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     3032..3051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3270..3374)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3405..3784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3786..4043
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4090..4215
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4707..4824
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5184..5417
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5602..5646
                     /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        5903..6206
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6207..6410
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6361..6381
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6486..6495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6492..6494,6495..6497,6498..7208)
                     /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(6537..6558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6798..6817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7145..7166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaagt ggctccgcgt ccgtcaatac cggtgaaaca aaaaccaaaa gaaaaagaga
       61 aacccccccc ggttaataag caagaaaacg caggcacgct gaacatcctg tccacgctca
      121 gtaatggtaa ctcgtctaag cagaaaatcc cggctgatgg cgttcatcgt tgaaccggtt
      181 ctagataaga ttacggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
      241 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag aagacaagat
      301 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
      361 ggctaactag ggaacccact gcttaagcct caataaagct tcagctgctc gagctagcag
      421 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
      481 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
      541 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
      601 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
      661 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
      721 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
      781 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
      841 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
      901 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     1081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     1141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1201 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1261 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1381 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     1441 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2461 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2521 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2581 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2641 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2701 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2761 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2821 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2881 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2941 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3001 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3061 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3121 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3181 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3241 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3301 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3361 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac
     3421 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     3481 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3541 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3781 catgggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     3841 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     3901 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     3961 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4021 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     4081 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     4141 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     4201 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     4261 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     4321 ctagaacgat tcgcagttaa tactggcctg ttagaaacat cagaaggctg tagacaaata
     4381 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     4441 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     4501 ttagacaaga tagaggaaga gcaaaacaaa agtaagaaaa aagcacagca agcagcagga
     4561 tcttcagacc tggaaattcc ctacaatccc caaagtcaag gagtagtaga atctatgaat
     4621 aaagaattaa agaaaattat aggacaggta agagatcagg ctgaacatct taagacagca
     4681 gtacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     4741 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     4801 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaaa tccactttgg
     4861 aaaggaccag caaagctcct ctggaaaggt gaaggggcag tagtaataca agataatagt
     4921 gacataaaag tagtgccaag aagaaaagca aagatcatta gggattatgg aaaacagatg
     4981 gcaggtgatg attgtgtggc aagtagacag gatgaggatt agaacatgga aaagtttagt
     5041 aaaacaccat aaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     5101 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     5161 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     5221 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     5281 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     5341 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     5401 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     5461 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     5521 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     5581 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     5641 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     5701 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     5761 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccg
     5821 agctcaagct tcgaacgcgt gctagttatt aatagtaatc aattacgggg tcattagttc
     5881 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5941 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6001 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6061 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6121 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6181 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     6241 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6301 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6361 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     6421 accgggctcg tttagtgaac cgaattctgc agtcgacggt accgcgggcc cgggatccac
     6481 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     6541 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     6601 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     6661 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     6721 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     6781 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     6841 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     6901 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     6961 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     7021 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     7081 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     7141 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     7201 tgtacaaggg cggaagcgga ggctccgggg gaagtgcggc cgcttcccgg tggatgccgt
     7261 ccaaagcgta tctgcagaaa caggcgaaag ctgtgaagaa gaaggagaaa aagtcgaaaa
     7321 cctctgaaaa gaaggattcc aaggaaagct ctgtcgtgaa aaacgtcgtg gactcatccc
     7381 aaaagccgac tccgagcgcg cgtgaggacc cggcaccgaa aaagagctcc agcgagccgc
     7441 caccgcggaa accggttgaa gagaaatccg aggagggtaa cgtttcggcc ccgggtcccg
     7501 agagtaaaca agcgaccaca ccagcgagcc gcaaatccag caagcaggtc agccagccag
     7561 ctctcgttat tccgccgcag ccgccgacca ctgggccgcc gcgtaaggaa gtgccgaaaa
     7621 ccaccccgtc ggagccgaaa aagaagcagc cgccgccgcc ggagagcggt ccggaacaaa
     7681 gtaagc
//
