LOCUS       sequence-487993-        7683 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_086, 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 7683)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7683)
  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_086
FEATURES             Location/Qualifiers
     source          1..7683
                     /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(1216..1926,1927..1929,1930..1932))
                     /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(1258..1279)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1607..1626
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1866..1887
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1929..1938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2014..2217)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2043..2063)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2218..2521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2778..2822)
                     /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    3007..3240
                     /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    3600..3717
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4209..4334
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4381..4638
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4640..5019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5050..5154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5155..5223,5224..6015)
                     /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(5373..5392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6186..6774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6675..6694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6928..6945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7013..7147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7037..7056)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7091..7110
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7153..7348)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7167..7302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7407..7428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7443..7448,7449..7466,7467..7473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7481..7497
                     /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     7486..7508
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7505..7521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7505..7521
                     /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 gctggtccag cggaaggtcg gttcacggag
      601 aatggactta cgcttaccct gcataggagc ggttgacgcc ggcaaaaacg gagatgccaa
      661 cgggatcgtc ggcggcatca gccacggggt gtggtcgctg atagagctgg ccggctgcag
      721 tggcggcggc ggagttagca gcgggggcgg cggggaacta gaggtctgtt gttgcggcgc
      781 gctttgcagt gttgacagct ttttagtggt acgagagccg aagctacgct cggaaacgct
      841 gtagcggcga gagcgacgat cattgctttc attctccgga gattggctga ttggaagggg
      901 cggatgcacc tccggggtat cagaacgctc ctccggcagg acttggatct cctcgcttgc
      961 ctggctgtca gtactcgtat caacgctcgg ggatgaggaa cgagacgaat cagagctcat
     1021 ttgtgaggaa tgctgactcg cagcggaaga cttttcggag ctgccacagc tcggagcgct
     1081 aaagcggctg ttcggggtag attccagacg cgcgattttg atcggcggat cgtagtcttc
     1141 gtcttctatg aagcgacgag gggtcttgat tatccgggaa gcggccgcac ttcccccgga
     1201 gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
     1261 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
     1321 ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     1381 ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt
     1441 caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt
     1501 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     1561 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     1621 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     1681 gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag
     1741 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag
     1801 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     1861 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     1921 cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat
     1981 tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga cctcccaccg
     2041 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     2101 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     2161 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     2221 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     2281 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     2341 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     2401 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     2461 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     2521 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     2581 agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat
     2641 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     2701 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     2761 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     2821 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     2881 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     2941 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     3001 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     3061 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     3121 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     3181 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     3241 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     3301 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     3361 atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc atcctgtcta
     3421 cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat ctttgctttt
     3481 cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc ttcacctttc
     3541 cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta ataaacccga
     3601 aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt
     3661 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg
     3721 tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc tcttacctgt
     3781 cctataattt tctttaattc tttattcata gattctacta ctccttgact ttggggattg
     3841 tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt acttttgttt
     3901 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     3961 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     4021 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     4081 gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     4141 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     4201 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     4261 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     4321 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     4381 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     4441 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     4501 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     4561 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
     4621 ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4681 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4741 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4801 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4861 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4921 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4981 taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg cacttttcgg
     5041 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5161 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5221 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5281 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5341 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5401 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5461 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5521 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5581 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5641 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5701 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5761 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5821 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5881 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5941 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6001 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6301 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6781 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6841 cctgcgttat cccctgcttc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6901 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6961 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag cggatccaga
     7021 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     7081 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     7141 acaagttagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     7201 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     7261 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     7321 gactatggtt gctgactaat tgagatgcgg atccgctggc acgacaggtt tcccgactgg
     7381 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7441 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7501 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     7561 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     7621 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     7681 tat
//
