LOCUS       sequence-487999-        7365 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_094, derived from human fusion protein KMT2A_MLLT10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7365)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7365)
  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_094
FEATURES             Location/Qualifiers
     source          1..7365
                     /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(898..1608,1609..1611,1612..1614))
                     /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(940..961)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1289..1308
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1548..1569
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1611..1620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1696..1899)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1725..1745)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1900..2203
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2460..2504)
                     /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    2689..2922
                     /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    3282..3399
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3891..4016
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4063..4320
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4322..4701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4732..4836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4837..4905,4906..5697)
                     /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(5055..5074)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5868..6456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6357..6376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6610..6627
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6695..6829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6719..6738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6773..6792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6835..7030)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6849..6984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7089..7110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7125..7130,7131..7148,7149..7155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7163..7179
                     /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     7168..7190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7187..7203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7187..7203
                     /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 ggacttttct tgggcgaccg gaccggtttt
      601 gtctgacaag cgagcaactt tttgggatgc gttgtcaccg tgaatcgtca aaaacgggtt
      661 cgtggtcgcc gtgtgtaacg gcggcgcctg tgtacctgcc agcaaattgt tgatcggaat
      721 ctgcgttgga ccagggattt gtagctgctg cagctccggt tggtgatgct gctgatggtg
      781 gtgctgcatt aactggtaca gaaacgcttg gtgctgttcc ggcgtcaatt gctggctatt
      841 gagcagctgc tgaaaccggg aagcggccgc acttcccccg gagcctccgc ttccgccctt
      901 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      961 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt
     1021 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     1081 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     1141 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     1201 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1261 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     1321 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1381 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     1441 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     1501 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     1561 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     1621 gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca ctaaacgagc
     1681 ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     1741 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     1801 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     1861 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     1921 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     1981 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     2041 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2101 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     2161 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     2221 gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg ttcgaagctt
     2281 gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc
     2341 ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct
     2401 actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac
     2461 aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc
     2521 gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc
     2581 atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa
     2641 ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga
     2701 tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact
     2761 gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc
     2821 actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata
     2881 gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt
     2941 tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt
     3001 actactttat atttatataa ttcacttctc caattgtccc tcatatctcc tccttatggt
     3061 gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac acaatcatca
     3121 cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg cactactttt
     3181 atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag ctttgctggt
     3241 cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg aatttttgta
     3301 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     3361 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     3421 tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat tttctttaat
     3481 tctttattca tagattctac tactccttga ctttggggat tgtagggaat ttccaggtct
     3541 gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc ctctatcttg
     3601 tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata gagggttgct
     3661 actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg ttgtagctgt
     3721 cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt
     3781 tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt tccccctggc
     3841 cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact gacgctctcg
     3901 cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact caccagtcgc
     3961 cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc
     4021 tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag attttccaca
     4081 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     4141 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     4201 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag gcaaaaagca
     4261 gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag gccacgcctc
     4321 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     4381 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     4441 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     4501 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     4561 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     4621 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     4681 aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc
     4741 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4801 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4861 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4921 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4981 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5041 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5101 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5161 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5221 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5281 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5341 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5401 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5461 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5521 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5581 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5641 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5701 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5761 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5821 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5881 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5941 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6001 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6061 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6121 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6181 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6241 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6301 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6361 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6421 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6481 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgct
     6541 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6601 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6661 ctccccgcgc gttggccgat tcattaatgc agcggatcca gacatgataa gatacattga
     6721 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     6781 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta gctttttgca
     6841 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     6901 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     6961 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     7021 attgagatgc ggatccgctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7081 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7141 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7201 gaccatgatt acgccaagct tgggctgcag gtcgagggat ctccataaga gaagagggac
     7261 agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga
     7321 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatat
//
