LOCUS       sequence-488132-        7350 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_109, derived from human fusion protein MKRN1_BRAF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7350)
  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_109
FEATURES             Location/Qualifiers
     source          1..7350
                     /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(883..1593,1594..1596,1597..1599))
                     /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(925..946)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1274..1293
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1533..1554
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1596..1605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1681..1884)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1710..1730)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1885..2188
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2445..2489)
                     /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    2674..2907
                     /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    3267..3384
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3876..4001
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4048..4305
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4307..4686
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5853..6441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6342..6361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6595..6612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6680..6814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6704..6723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6758..6777
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6820..7015)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6834..6969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7074..7095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7110..7115,7116..7133,7134..7140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7148..7164
                     /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     7153..7175
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7172..7188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7172..7188
                     /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 tgtaatttgg ccgtcgggga tctcccagtc
      601 gtcagaacta tcgcggcgac caagggtctt catacgatta cgatcctcgg acgagctgct
      661 ggacttgcgc tcacgttgcg gtcccggcga cttttgaagg gctttcacat tggtcagcga
      721 tccaggtaaa ctagcggggg gtgttgcgga cagtcctgtg gtcgaaccgc catctccgcg
      781 aaagccctga tcacggatta agtccttgat gtgttgagaa cgctgcgcag catccattgg
      841 ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca gctcgtccat
      901 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      961 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
     1021 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1081 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     1141 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1201 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1261 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1321 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1381 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1441 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1501 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1561 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
     1621 ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt tcactaaacg
     1681 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     1741 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     1801 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     1861 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     1921 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     1981 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     2041 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     2101 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     2161 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     2221 ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct cggtcccctc
     2281 ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac tctattcact
     2341 atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat cattatgaat
     2401 aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact tgcccattta
     2461 tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc ttcaattaag
     2521 gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca ggtcgtgtga
     2581 ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac agcagtggtg
     2641 caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt taggtatctt
     2701 tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag ttgcaacaga
     2761 tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat accagacaat
     2821 aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct
     2881 cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct ctgcaccact
     2941 cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac tttatattta
     3001 tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt actaaacttt
     3061 tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc catctgtttt
     3121 ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc actattatct
     3181 tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt ccaaagtgga
     3241 tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     3301 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     3361 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac tgctgtctta
     3421 agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt attcatagat
     3481 tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga tcctgctgct
     3541 tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     3601 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     3661 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     3721 cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag ctccctgctt
     3781 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     3841 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     3901 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     3961 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     4021 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     4081 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     4141 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     4201 ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc ttatatgcag
     4261 gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg gtaatagcga
     4321 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4381 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4441 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4501 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4561 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4621 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     4681 aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5881 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5941 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6001 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6061 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgcttctgt ggataaccgt
     6541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6601 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6661 ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt ttggacaaac
     6721 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6781 atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc ctaggcctcc
     6841 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     6901 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6961 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcggatc
     7021 cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7081 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7141 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7201 aagcttgggc tgcaggtcga gggatctcca taagagaaga gggacagcta tgactgggag
     7261 tagtcaggag aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt
     7321 taaagaaaaa aataacacaa aacaaaatat
//
