LOCUS       sequence-487994-        8607 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_087, 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 8607)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8607)
  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_087
FEATURES             Location/Qualifiers
     source          1..8607
                     /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(2140..2850,2851..2853,2854..2856))
                     /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(2182..2203)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2531..2550
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2790..2811
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2853..2862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2938..3141)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2967..2987)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3142..3445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3702..3746)
                     /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    3931..4164
                     /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    4524..4641
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5133..5258
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5305..5562
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5564..5943
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5974..6078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6079..6147,6148..6939)
                     /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(6297..6316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7110..7698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7599..7618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7852..7869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7937..8071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7961..7980)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8015..8034
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8077..8272)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8091..8226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8331..8352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8367..8372,8373..8390,8391..8397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8405..8421
                     /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     8410..8432
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8429..8445
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8429..8445
                     /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 acaacgacga gaacggcggc ccttcttaac
      601 aggcggctct tgcggcgctt tattacgggt caccggttta atcggcttaa tcggcggagc
      661 cagcggctca gcatcctcgc tacccgcgat cgatgactta tcatcgttac ccatgctgct
      721 cagaatcttc tcgcgttctt cccacggcag tgcgctcagc gtcggcatgt cgtctgggaa
      781 aactgcgcgc ttacggccca gcgcaacggc agcacgacgg caaacgtgtt taatgcgcgg
      841 acctctcacc gatgtctcac tactatcact ttcttgacct tgagcttttg gctgatccgt
      901 ttgtttgagc gacttcgatt tttcaatttt gcacagctga gcctttgcct tcttgagcag
      961 gctcgcaaca cgcttatcgg tcataggcaa tttgtccgct tgagccagca tagagccgat
     1021 cgaggaggtg gagtgcttca cggtagagga gctcggcgtg ctcaggcaca aggttttctc
     1081 cttctccaga ctcggagccg tcgggcccaa atccaggttg gtcttctcca agttgccacg
     1141 gccctttttg attaagattt tggtcttcac cgcagtcgta tcacccaggg tgacgctggt
     1201 gatatccgtt ccggagtcgt gagagctcga tttcttacga ccggtcgcct tcttggcgga
     1261 agaggaggta gcaacgtctt caccaaccac tttctcttta gacacacggc caaccggata
     1321 cagcgcagag ctgctctgga tttcgctacc tttctttcgt ttttctttac gagattccct
     1381 cttgttctct ttttcgcgct cacgatcacg ctcacggctc ttgtcttttt ccacgctctt
     1441 gtcagcatcc cggtccttgg atagctcctc cggcgctttg tctttgttac gaccgcgctc
     1501 ggtctgactg cccggggtaa accacgggaa cagcggcgtt ggggaggagg agctaaacgg
     1561 ttcagctggc gcgctcgtct gcttacgcgg acgctggttc ttctccgcgc tttcgccact
     1621 ctgcgtcagc gagtgagacg gaaaggtgaa ggtcgggttc aatgcactgg tcgcgagcgg
     1681 gctcacagag atactgaggc tactgctcac ggaggaaggc ggggtcagag gtgacagttc
     1741 actggagctc aggcgaccgc tacgcgttct catggagtgc gacgggctac gaggctccga
     1801 acgaatcggg ctgaacactt ttcgctttct cttacgatta gacacaccag agctgctagt
     1861 acctgcgctg gtacggttag atgggagggt tacgctttca aagatacggc tgtgtgcctc
     1921 agacggggtg aagcgcgggg cacgtaacag cggtgaacgc ttatgcatat cgaaacgggt
     1981 cccggaatgc agcggactaa agagacgggc agacgctgca gtgcctgatg cagaaaagcc
     2041 ggacgcgaag cccacgtctt ccggagtcaa tggcggcgga cggaaattat caaagatccg
     2101 ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct cgtccatgcc
     2161 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     2221 ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     2281 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     2341 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     2401 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     2461 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     2521 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     2581 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     2641 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     2701 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     2761 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     2821 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg gatcccgggc
     2881 ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca ctaaacgagc
     2941 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     3001 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     3061 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     3121 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     3181 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     3241 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     3301 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     3361 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     3421 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     3481 cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg tcccctcggg
     3541 gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata
     3601 gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat
     3661 tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct
     3721 aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag
     3781 tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc
     3841 caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa
     3901 atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc
     3961 acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc
     4021 tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat
     4081 tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc
     4141 aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt
     4201 ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat
     4261 aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact aaacttttcc
     4321 atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat ctgttttcca
     4381 taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact attatcttgt
     4441 attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca aagtggattt
     4501 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     4561 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     4621 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc tgtcttaaga
     4681 tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt catagattct
     4741 actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc tgctgcttgc
     4801 tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
     4861 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
     4921 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
     4981 ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc cctgcttgcc
     5041 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
     5101 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
     5161 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
     5221 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
     5281 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
     5341 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
     5401 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
     5461 agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat
     5521 ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta atagcgatga
     5581 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     5641 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     5701 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     5761 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     5821 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     5881 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     5941 gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6001 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6061 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6121 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6181 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6241 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6301 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6361 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6421 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6481 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6541 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6601 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6661 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6721 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6781 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6841 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6901 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6961 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7021 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7081 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg cttctgtgga taaccgtatt
     7801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7861 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7921 attcattaat gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     7981 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     8041 tgtaaccatt ataagctgca ataaacaagt tagctttttg caaaagccta ggcctccaaa
     8101 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     8161 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     8221 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcggatccgc
     8281 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8341 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8401 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8461 cttgggctgc aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag
     8521 tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa
     8581 agaaaaaaat aacacaaaac aaaatat
//
