LOCUS       sequence-487998-        8229 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_092, derived from human fusion protein KMT2A_MLLT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8229)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8229)
  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_092
FEATURES             Location/Qualifiers
     source          1..8229
                     /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(1762..2472,2473..2475,2476..2478))
                     /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(1804..1825)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2153..2172
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2412..2433
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2475..2484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2560..2763)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2589..2609)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2764..3067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3324..3368)
                     /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    3553..3786
                     /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    4146..4263
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4755..4880
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4927..5184
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5186..5565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5596..5700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5701..5769,5770..6561)
                     /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(5919..5938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6732..7320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7221..7240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7474..7491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7559..7693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7583..7602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7637..7656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7699..7894)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7713..7848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7953..7974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7989..7994,7995..8012,8013..8019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8027..8043
                     /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     8032..8054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8051..8067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8051..8067
                     /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 aatcagattc acaatctgtt gcagcacgtt
      601 gcgctctctc agtgccataa ggcgacggtg caattcaacc aactcgtcgg tgtacgcttt
      661 gtcgtacgtc cctttcttta agatcttctc cggcttgctg cagctctcag ggctacgacg
      721 gccgctcacc ttgctatttg gcggcggcgg cttctgaggc ggaggtggct cacgagatgg
      781 caggctactg tccgcggagt tgtcgctttc actatccgag aacgagagcc tgctgtcacg
      841 acccgggttg gttttgcccg cggcctcctc gcctgagctc gaatcgtctt catcggactc
      901 ctcactttgc aggtcctcta ccatgcttct caacgggccc tggctgtggt tttgcgaggg
      961 ttcaaagtca gagtccgacg agctgtcgct agaactgcta gagttgctcg gtgatgattg
     1021 cgcagattct gatttgaagc tcgcttcatc ttcgctatta ctctcttcaa cctctaaagc
     1081 ctttttcgcc tcacgcggct cggattctgc cttcactttc tcaccacggg tgctagactt
     1141 gtccttagcc ggctttttat cagagaatga actgctagaa gaagtacgag gagatgtacc
     1201 tggggcggaa cgtgagcctt tggaagagct ctttttctgc ttcttcgcac tcggtttcgg
     1261 ggagtcagcc gtcgccggcc gctttgagga tgcgcgtggc ggcggtggcg gcggtggcgg
     1321 cggtccgcct tttggcgaag ttgattccag tttggtctct ttcagggcca ttttgggctc
     1381 cttaaacgcc gccttgggcg gcggtgcctt ttcttctttc ggcaatctac cctcaccaag
     1441 tttacgactg gtatccttgg agcttttcgc ttgttccctc tccagctctt tcgaagacga
     1501 tttcgactcg gaatctttcc gaggacgttc gcgatgttct ttggtaactt tatgcggttt
     1561 tgaggtcttc gagctttctt tatttgcatc tttgctgccg tgtgacggct tggtcttttt
     1621 cgggtcagag aacgcactca gcgggatcgt cggcagcatc gggtaatcgg ggctcggacg
     1681 ggacacggta tcggcaccct ccggcataac catgacaccc cgggaagcgg ccgcacttcc
     1741 cccggagcct ccgcttccgc ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1801 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1861 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1921 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     1981 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     2041 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     2101 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     2161 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     2221 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     2281 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     2341 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     2401 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     2461 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg
     2521 cagaattcgg ttcactaaac gagcccggtt cactaaacga gctctgctta tatagacctc
     2581 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2641 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2701 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     2761 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     2821 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     2881 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     2941 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     3001 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     3061 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     3121 ataactagca cgcgttcgaa gcttgagctc ggtcccctcg gggttgggag gtgggtctga
     3181 aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact
     3241 attcttaaac ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa
     3301 tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt
     3361 tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa
     3421 ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat
     3481 ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa
     3541 ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc
     3601 tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc
     3661 ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc
     3721 agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa
     3781 gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt
     3841 gctactccta atggttcaat ttttactact ttatatttat ataattcact tctccaattg
     3901 tccctcatat ctcctcctta tggtgtttta ctaaactttt ccatgttcta atcctcatcc
     3961 tgtctacttg ccacacaatc atcacctgcc atctgttttc cataatccct aatgatcttt
     4021 gcttttcttc ttggcactac ttttatgtca ctattatctt gtattactac tgccccttca
     4081 cctttccaga ggagctttgc tggtcctttc caaagtggat ttctgctgtc cctgtaataa
     4141 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     4201 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     4261 aaattgtgga tgaatactgc catttgtact gctgtcttaa gatgttcagc ctgatctctt
     4321 acctgtccta taattttctt taattcttta ttcatagatt ctactactcc ttgactttgg
     4381 ggattgtagg gaatttccag gtctgaagat cctgctgctt gctgtgcttt tttcttactt
     4441 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     4501 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     4561 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     4621 aggccagtat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     4681 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     4741 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     4801 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     4861 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     4921 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     4981 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     5041 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     5101 gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc
     5161 ccagtcccgc ccaggccacg cctcccatgg taatagcgat gactaatacg tagatgtact
     5221 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     5281 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     5341 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     5401 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     5461 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     5521 accccgtaat tgattactat taataactag tcaataatca atgtcgaccc aggtggcact
     5581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5641 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5701 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5761 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5821 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5881 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5941 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6001 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6061 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6121 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6181 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6241 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6301 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6361 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6421 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6481 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6541 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6601 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6661 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6721 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6781 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6841 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6901 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6961 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7021 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7081 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7141 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7201 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7261 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7321 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7381 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7441 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7501 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcgga
     7561 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     7621 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     7681 caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     7741 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     7801 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     7861 gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga caggtttccc
     7921 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7981 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8041 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgag
     8101 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
     8161 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
     8221 acaaaatat
//
