LOCUS       sequence-487992-        9252 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_085, derived from human fusion protein KMT2A_AFDN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9252)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 9252)
  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_085
FEATURES             Location/Qualifiers
     source          1..9252
                     /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(2785..3495,3496..3498,3499..3501))
                     /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(2827..2848)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3176..3195
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3435..3456
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3498..3507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3583..3786)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3612..3632)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3787..4090
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4347..4391)
                     /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    4576..4809
                     /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    5169..5286
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5778..5903
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5950..6207
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6209..6588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6619..6723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6724..6792,6793..7584)
                     /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(6942..6961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7755..8343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8244..8263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8497..8514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8582..8716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8606..8625)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8660..8679
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8722..8917)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8736..8871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8976..8997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9012..9017,9018..9035,9036..9042)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9050..9066
                     /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     9055..9077
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9074..9090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9074..9090
                     /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 cttggtgttc agttcgttct ccagttccgt
      601 tagcttacgg ctggccttaa ctttgttact cacgtcttga ccctgagaga agaggcgctg
      661 gcgttctttg aacgtcaggt tttccggcgc gccgctagag cccgggctat cagcatcctg
      721 gctttttgaa cgtttttcct ttgcgtcacg gcaggcatca tgtgcaccca cagctgcgcc
      781 cgtgctaccc ggatagctgt tagggccagc caagctacag tcctcctcct cttcctcttc
      841 gttatacgcg acgaatttgg cggtgaagag ggaatccggc gacagaacct gggttttcag
      901 gtagcttgca ttgcgctgcg gcggcggcgg tggcgcaccg ggtgccgggc tcgggcttgg
      961 cggttcgtag tcacgcggca gaggcggacg gcataaaccc ggtgcttccg gctccaacaa
     1021 acggcgggcc gcctcatcgt gctgcctgcg acgttccgct tccaggcgac tataataacc
     1081 ctcttcttga cgccgttttt cctccgcgtc gcgcttggta cgctcctcct cctgccgcct
     1141 acgctcttct tcttgacgcg cgcggtcctc cgcttcgcgt ttacgcattt cttccaattg
     1201 ttgttgcctg cgcctttcct catcttggag acgtgcccga cgctctgcct ccagacgctg
     1261 cataatcagc atggtatcca catcatcatc ctcctcttct tcgtcatctt cgtccttttg
     1321 cttgctctcc tgtaagcgtt tctggaactg ccattcaagc atcagcttac gcagacggtc
     1381 tgactcctcc gcgctacgat ccggtttgct ttgcaactcc tggatctcct ttgacagcat
     1441 atcaacgatg tgcatttgtt gctgcttctc cagtttctcc ttcgcatcgc gtttccacgg
     1501 gtccggggac aggctatcgc cagaggacag ttcctccttg gacacggtaa tatattgcag
     1561 atcacgacgt tcgatggtgc gcggttgttg ctgcggctgc agctcgcgga tcacggtttc
     1621 ctgcggtcgt tgcaacgtgg acggcttttc tggtttcatc tgctgcgcag tcaacgggga
     1681 aaacggagcc gggttcaggc tctgggtacg catctggccc agtttgcgct cttgttcgcg
     1741 gcgcttgcgc tcacgctctt cttcaagccg cgccttctct ttttcatacc aacgctggtg
     1801 ctcctcacgc tttctccgct ctgccgcggc aacctgggcg ctaggcagac caatttggtt
     1861 cgcgctcggc ggcggcggca ggggcaaatc catgctcata ccatcgaagt cacctgcata
     1921 atgtaccggc ggcggtggcg gcggcggcgg cagatcggta cgaatcggct gggaaacagc
     1981 aaccggggtc gctggaatcg cagccggggt tttccatcgg cccgggccgg acttggtcag
     2041 ggtggaggca ggagtgacgc ttttgctgct gtggttcagg tgctcttggc tgctcgtgga
     2101 ggagtccagg gaactcgact gatttatcca catatcgctg tcgctcttac gatccatagc
     2161 agcctcgata cgatgacgct ccagctggta cgctttgtcc tcgcgtaatt cctcctgcga
     2221 acgggtcaca cgttgaatgg caatagagct gtgattgctg tcggtatgca tgtgcggttt
     2281 ttcttcatag ttcggccact gattctgcgg cggcgccata cgatcttggc tcttggaggc
     2341 cggaaaggtg aaatattcac gggtataggt ttgcgtcggg atcgggtatg cctccggacg
     2401 tggtggcggg gtctgttctt cggtacacag gtttccggtg ctgacgctgg tgattttggc
     2461 tgtggtgccg ctcgcgtacg cggacttacc acccggagac gggggttgat tcgccacgtt
     2521 tggagaagaa cggtgatccg ctctgttctt catcagacgg tcatcccccg gcagcttctt
     2581 cggctcgctg tactctgccc aaggcagctg cggggattcc gggctaccgt tctgggtaga
     2641 gttattatag agctcaaagc cctcgctctt cggacgcggc ttaccggaac cgcggcgatc
     2701 gctgatacgc tgcatcatgg gagagggctg gttcagaagg gtccgggaag cggccgcact
     2761 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
     2821 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     2881 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     2941 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     3001 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     3061 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     3121 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     3181 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     3241 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     3301 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     3361 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     3421 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     3481 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     3541 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     3601 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     3661 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     3721 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     3781 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     3841 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     3901 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3961 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     4021 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     4081 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     4141 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     4201 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     4261 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     4321 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     4381 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     4441 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     4501 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     4561 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     4621 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     4681 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     4741 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     4801 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     4861 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     4921 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     4981 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     5041 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     5101 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     5161 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     5221 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     5281 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     5341 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     5401 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     5461 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     5521 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     5581 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     5641 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     5701 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     5761 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     5821 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     5881 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     5941 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     6001 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     6061 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     6121 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     6181 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     6241 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     6301 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     6361 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     6421 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     6481 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     6541 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     6601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6721 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6781 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6841 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6901 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6961 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7021 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7081 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7141 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7201 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7261 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7321 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7381 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7441 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7501 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7561 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7861 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8401 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8461 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8521 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8581 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     8641 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     8701 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     8761 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     8821 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     8881 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     8941 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     9001 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     9061 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     9121 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     9181 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     9241 aaaacaaaat at
//
