LOCUS       sequence-487968-        7785 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_055, derived from human fusion protein EML4_BRAF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7785)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7785)
  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_055
FEATURES             Location/Qualifiers
     source          1..7785
                     /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(1318..2028,2029..2031,2032..2034))
                     /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(1360..1381)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1709..1728
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1968..1989
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2031..2040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2116..2319)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2145..2165)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2320..2623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2880..2924)
                     /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    3109..3342
                     /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    3702..3819
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4311..4436
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4483..4740
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4742..5121
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5152..5256
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5257..5325,5326..6117)
                     /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(5475..5494)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6288..6876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6777..6796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7030..7047
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7115..7249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7139..7158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7193..7212
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7255..7450)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7269..7404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7509..7530
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7545..7550,7551..7568,7569..7575)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7583..7599
                     /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     7588..7610
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7607..7623
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7607..7623
                     /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 ggtgatctgc ccatcgggga tctcccagtc
      601 gtcagaggag tcacgacgcc ccagggtctt catacggtta cggtcttcgc tagacgagct
      661 gcttttacgc tcgcgttgtg ggccgggtga cttctgaagt gcctttacat tggtcagcga
      721 accaggcaga ctggcaggcg gggtcgccga cagacccgtt gttgatcctt ggttgatgat
      781 cacatcctta tgtttgtccg ccgtctttgt aactttcggg ataagtttcg gagtgctggg
      841 aatcttcgat aacttgttgc gggaatcatc actattttcc caggagttat ggcttttttc
      901 ggctgggcta ggacgcttaa tactcttagt cggggtggcg tttttgcttt ctggcgtctg
      961 gcgatggatt tgtaacggct gactgcttgg ctgcggagaa ggcgaggcac ggatctgggg
     1021 tgactgatcg ttcgagtgcg attcctcctt cttttcgcgt tgcccttgcg gtttttcctt
     1081 cttcttctcg gttcctgatt ttgctgcaga ggataatgtc tctttgccag caatggaaac
     1141 cgcactcgta tgagagggct tacgattagc gccggagccg ttcgtgatgc agctcatcgg
     1201 aataaccgcg cggggagagg gttgaccctt tgatgatacc gatttcttga ctgacgctac
     1261 gtgatcctca ctgatccggg aagcggccgc acttcccccg gagcctccgc ttccgccctt
     1321 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     1381 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt
     1441 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     1501 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     1561 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     1621 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1681 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     1741 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1801 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     1861 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     1921 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     1981 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     2041 gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca ctaaacgagc
     2101 ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     2161 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     2221 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     2281 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     2341 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     2401 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     2461 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2521 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     2581 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     2641 gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg ttcgaagctt
     2701 gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc
     2761 ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct
     2821 actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac
     2881 aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc
     2941 gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc
     3001 atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa
     3061 ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga
     3121 tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact
     3181 gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc
     3241 actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata
     3301 gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt
     3361 tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt
     3421 actactttat atttatataa ttcacttctc caattgtccc tcatatctcc tccttatggt
     3481 gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac acaatcatca
     3541 cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg cactactttt
     3601 atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag ctttgctggt
     3661 cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg aatttttgta
     3721 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     3781 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     3841 tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat tttctttaat
     3901 tctttattca tagattctac tactccttga ctttggggat tgtagggaat ttccaggtct
     3961 gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc ctctatcttg
     4021 tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata gagggttgct
     4081 actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg ttgtagctgt
     4141 cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt
     4201 tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt tccccctggc
     4261 cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact gacgctctcg
     4321 cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact caccagtcgc
     4381 cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc
     4441 tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag attttccaca
     4501 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     4561 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     4621 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag gcaaaaagca
     4681 gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag gccacgcctc
     4741 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     4801 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     4861 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     4921 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     4981 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     5041 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     5101 aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc
     5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5221 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5281 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5341 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5401 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5461 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5521 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5581 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5641 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5701 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5761 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5821 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5881 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5941 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6001 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6061 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6121 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgct
     6961 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7021 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7081 ctccccgcgc gttggccgat tcattaatgc agcggatcca gacatgataa gatacattga
     7141 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     7201 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta gctttttgca
     7261 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     7321 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     7381 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     7441 attgagatgc ggatccgctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7501 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7561 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7621 gaccatgatt acgccaagct tgggctgcag gtcgagggat ctccataaga gaagagggac
     7681 agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga
     7741 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatat
//
