LOCUS       sequence-490724-        7473 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_082, derived from human fusion protein KMT2A_AFDN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7473)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7473)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_082
FEATURES             Location/Qualifiers
     source          1..7473
                     /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(1006..1716,1717..1719,1720..1722))
                     /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(1048..1069)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1397..1416
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1656..1677
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1719..1728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1804..2007)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1833..1853)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2008..2311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2568..2612)
                     /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    2797..3030
                     /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    3390..3507
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3999..4124
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4171..4428
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4430..4809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4840..4944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4945..5013,5014..5805)
                     /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(5163..5182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6718..6735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6803..6937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6827..6846)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6881..6900
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6943..7138)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6957..7092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7197..7218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7233..7238,7239..7256,7257..7263)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7271..7287
                     /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     7276..7298
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7295..7311
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7295..7311
                     /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 tgcatagatt cgaagggtac cgccactatc
      601 gggacggccg tcactgctgc ggaactcttg catacgcttt tccagttttt gttgccgacg
      661 acgtttcatg accacctccg ggttgctgat ggtacgggta aagctggtct ccggcatatc
      721 tttgtacact tccgccgcca gccgactgtt ttcgacgtcc tcaccctgaa atggacggtc
      781 gtccttgtcg gacgcctgcc taagcgcttc tttttcgcgc ttcttctttt cctttttttc
      841 ctttttgctc aaggtacgct tgaaattctg aataacgcct tccttctctt gcttctcagg
      901 gccgttggac tgcgcttttt tcggcggaat ggcatcgttc tcgttcttca gaacaaaacg
      961 accccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
     1021 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1081 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     1141 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1201 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1261 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1321 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1381 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1441 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1501 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1561 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1621 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1681 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     1741 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     1801 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     1861 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     1921 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     1981 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     2041 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     2101 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     2161 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2221 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2281 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2341 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2401 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2461 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2521 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2581 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     2641 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     2701 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     2761 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     2821 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     2881 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     2941 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     3001 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     3061 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     3121 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3181 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3241 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3301 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3361 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3421 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3481 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3541 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3601 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     3661 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     3721 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     3781 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     3841 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     3901 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     3961 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     4021 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     4081 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     4141 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4201 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4261 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4321 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4381 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4441 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4501 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4561 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4621 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4681 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4741 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     4801 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4921 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4981 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5041 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5101 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5161 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5221 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5281 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5341 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5401 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5461 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5521 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5581 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5641 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5701 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5761 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6721 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6781 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     6841 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6901 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     6961 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     7021 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     7081 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     7141 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7201 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7261 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7321 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7381 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7441 tgttaaagaa aaaaataaca caaaacaaaa tat
//
