LOCUS       sequence-487935-        7683 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_020, derived from human fusion protein BRD9_CEP72.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7683)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7683)
  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_020
FEATURES             Location/Qualifiers
     source          1..7683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(651..670)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    716..771
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(770..789)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1132..1148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1132..1148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1145..1167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1156..1172
                     /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)."
     promoter        complement(join(1180..1186,1187..1204,1205..1210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1225..1246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1305..1500
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1351..1486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1506..1640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1543..1562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1597..1616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1708..1725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1879..2467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1959..1978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2638..3429,3430..3498))
                     /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     3261..3280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3499..3603)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3634..4013
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             4015..4272
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4319..4444
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4936..5053
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5413..5646
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5831..5875
                     /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"
     enhancer        6132..6435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6436..6639
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6590..6610
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6715..6724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6721..6723,6724..6726,6727..7437)
                     /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(6766..6787)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7027..7046)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7374..7395
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaaga agaaaagcga aaaggagaaa caccttgacg atgaagagcg tcgcaagcgt
       61 aaggaggaga agaagcgcaa acgtgagcgc gaacattgcg atacagaagg cgaggcggac
      121 gatttcgatc ctgggaagaa ggtcgaagta gagccacccc cagatcgtcc tgtacgtgct
      181 tgtcgcacac aacccggttt acaaactagc gtcaagcgcc tttgcggtga aatcgtagaa
      241 cttaagcaac atttagagca ttacgataaa atccaggaac tgacccaaat gttgcaggag
      301 tcacactcct cactggtatc aacaaacgag catttgctgc aagaactttc ccaagtgcgt
      361 gctcaacacc gtgccgaagt agagcaaatg cactggtcgt gaaccggttc tagataagat
      421 tacggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
      481 agaaaagggg ggactggaag ggctaattca ctcccaaaga agacaagatc tgctttttgc
      541 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      601 gaacccactg cttaagcctc aataaagctt cagctgctcg agctagcaga tctttttccc
      661 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
      721 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
      781 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
      841 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
      901 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
      961 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     1021 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     1081 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     1141 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1201 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1261 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     1321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     1381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     1441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1501 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1621 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaagcagg
     1801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3601 gcgcacattt ccccgaaaag tgccacctgg gtcgacattg attattgact agttattaat
     3661 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3721 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3781 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3841 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     3901 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     3961 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgggaggcg
     4021 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
     4081 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     4141 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     4201 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     4261 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     4321 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     4381 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     4441 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     4501 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     4561 cgcagttaat actggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     4621 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     4681 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     4741 agaggaagag caaaacaaaa gtaagaaaaa agcacagcaa gcagcaggat cttcagacct
     4801 ggaaattccc tacaatcccc aaagtcaagg agtagtagaa tctatgaata aagaattaaa
     4861 gaaaattata ggacaggtaa gagatcaggc tgaacatctt aagacagcag tacaaatggc
     4921 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     4981 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     5041 aattcaaaat tttcgggttt attacaggga cagcagaaat ccactttgga aaggaccagc
     5101 aaagctcctc tggaaaggtg aaggggcagt agtaatacaa gataatagtg acataaaagt
     5161 agtgccaaga agaaaagcaa agatcattag ggattatgga aaacagatgg caggtgatga
     5221 ttgtgtggca agtagacagg atgaggatta gaacatggaa aagtttagta aaacaccata
     5281 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     5341 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     5401 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     5461 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     5521 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     5581 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     5641 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     5701 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     5761 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     5821 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     5881 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     5941 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     6001 atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccga gctcaagctt
     6061 cgaacgcgtg ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     6121 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6181 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6241 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6301 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6361 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6421 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     6481 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     6541 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     6601 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgggctcgt
     6661 ttagtgaacc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc
     6721 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     6781 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     6841 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     6901 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     6961 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     7021 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     7081 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     7141 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     7201 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     7261 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     7321 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     7381 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     7441 ggaagcggag gctccggggg aagtgcggcc gcttcccgga tgggaaagaa acacaagaaa
     7501 cataaagcag aatggcgctc cagttacgaa gactacgctg ataaaccgtt agagaaacca
     7561 ttgaaattgg tattgaaagt tggaggttcg gaggtgacgg agttgtctgg ctctgggcat
     7621 gattcaagtt actacgacga ccgctccgat cacgagcgtg aacgtcacaa ggagaagaaa
     7681 aag
//
