LOCUS       sequence-488224-        9264 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_209, derived from human fusion protein YAP1_MAMLD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9264)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 9264)
  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_209
FEATURES             Location/Qualifiers
     source          1..9264
                     /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(2797..3507,3508..3510,3511..3513))
                     /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(2839..2860)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3188..3207
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3447..3468
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3510..3519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3595..3798)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3624..3644)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3799..4102
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4359..4403)
                     /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    4588..4821
                     /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    5181..5298
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5790..5915
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5962..6219
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6221..6600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6631..6735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6736..6804,6805..7596)
                     /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(6954..6973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7767..8355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8256..8275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8509..8526
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8594..8728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8618..8637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8672..8691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8734..8929)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8748..8883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8988..9009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9024..9029,9030..9047,9048..9054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9062..9078
                     /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     9067..9089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9086..9102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9086..9102
                     /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 aagagctggc atagaatcgc tcgcgacact
      601 acgctgaaag cgctgaggtt gttgcatcat ctgctgaagt aaaaatgacg agtgatccgg
      661 ctgctgctgc tgctgttgct gttgctgctg taactgtaag gtcgctgtag cagtggatga
      721 agccgtggcc gaggaagcct gggtaggtgt aggttgttgc aggtagtgca gtaaggacgg
      781 ttgacgcgac gttgtcggca tcgaaggcat tttttgaggt cccggttcgc tgacgaagtg
      841 agataaaggt ttcgtgtttc cgaacgtgaa aggttcggtg gcgccgggcg atgaagatgc
      901 catgccctgt tgcatgatca tgctcaaggt cttactggag tttgaggaga ttggcttgaa
      961 gattacgtta gcctgttgct gctgttgttg ctgttgctgt tgttgcgaca gcaggttcga
     1021 agtaggagta agtgaacgaa ttaaactaca ttgtggggtg aacgattgct gtggcaatcc
     1081 cggacttgac agcttttcag ggcggtacgg tgggctcggc ccaggattag atgcaggaag
     1141 ggctgacatc aggtggcctt gaggtgtttt aattgtgctg ctcatcaggt tggctaagat
     1201 agaagattgc ggcccaaatt ggggctgttg cgttaaggcg ggactaggca acttttcagg
     1261 agcgtagggc atcgctggac ccaaagctgc attgctgctg gatgtcatgg acgaaagcag
     1321 ggcattagga gacccgcgta aagtggagcc actcagcgtg ttagaactca tacaactgac
     1381 catcagactc tgtgggctga agggtggtgg tggcggggga aggggcaaag ggggcgggtg
     1441 cgggctggga acgggacgat aggggggtga caaacctggt ggcggcagac cagaccagct
     1501 cggagttggt ccttgttgct tagtcgctga cccctgtttg ctggctgcta atgcttttaa
     1561 ctgatgtgca tgatgccatt ggggaacggg cagaggcggc aaagcgacgg gcagcatggc
     1621 ttgaacttgc gacttggatt gggccatact cgatgacgga gagacaatct gcttggatgt
     1681 ggagggaatg ctgtaactaa tccccgtgga cgaactcgga atctgaaggc tcattccggt
     1741 cacttgactg cagctggaag caaattcttt tgaggaagct aaagattcta agtggctcat
     1801 ctggacggac ggcttaggcg tggtagacag cgttgcctgt ggatggtcaa gaaccagggg
     1861 ctcttctggt tttgtgccaa gaatcttttc aagatctaat tcattcggcg aagggtcctg
     1921 gatctttgtc agctcttcca gcaactcctg taattcctga tcgacagcgg gcacactatt
     1981 gattttctca taatagggta cagtagggcc cttcagtcca acttccgtgg tttgtggaac
     2041 tacgaaaggt gaacccataa tgttcccgtt catcggattg ttctccagca gaagggactg
     2101 ccccgccaca cccatagctg ctggagaagg attaatcgtc agcggaggca cctgaagttc
     2161 agcacatgca gttgatgggt gcgcgcccgg ccccatagcc agagttacgt cctccaagca
     2221 agggcgttta atcttattgg ggtatcctcc atccgccatg tccggaaatt ggaagtgatc
     2281 ttcttcttga cgacgcttca ccgtcccttg gcgcagcagc tcttgctgtt tcaagcgtaa
     2341 gcgttctttc tccatttgca attgctgcaa acgcatttgt tgctgctggt tcgagttgct
     2401 gccccccatt actccccctt gtggagactg cggtgcaagg ggcggaggtt gtttcacggg
     2461 tgccgactga gaaatacgct gattcatagc gaagcgcggg tccaggcgag ggtcaagcca
     2521 actcgtggtt ttgtttttat gattgatgta atagatctcg ccatcctggg tcattgcctg
     2581 ctcccagccg tcgggtaatg ggccagaagc cgagttcatc atgttctgct gcacgggcgg
     2641 ggatgtagga gctgtgacat tcatctggga cagcattgcc ttacgtgggt cttgccaggt
     2701 cgtcgtttga tcgatatggt ttaaaaagta gcgctgcccc gatgacgtct tagcccggga
     2761 agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt ccatgccgag
     2821 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     2881 gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     2941 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     3001 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     3061 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     3121 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     3181 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     3241 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     3301 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     3361 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     3421 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     3481 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg
     3541 cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta aacgagctct
     3601 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     3661 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     3721 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     3781 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     3841 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     3901 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     3961 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     4021 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     4081 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     4141 aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt
     4201 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
     4261 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
     4321 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
     4381 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
     4441 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     4501 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     4561 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     4621 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     4681 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     4741 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     4801 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     4861 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     4921 tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa cttttccatg
     4981 ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg ttttccataa
     5041 tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt atcttgtatt
     5101 actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag tggatttctg
     5161 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     5221 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     5281 cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt cttaagatgt
     5341 tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat agattctact
     5401 actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc tgcttgctgt
     5461 gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct
     5521 tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta
     5581 agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct
     5641 tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct gcttgcccat
     5701 actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat
     5761 cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag
     5821 cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg
     5881 cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc
     5941 aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     6001 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     6061 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     6121 tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat gcaggatctg
     6181 agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata gcgatgacta
     6241 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     6301 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     6361 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     6421 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     6481 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     6541 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     6601 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6961 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7261 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7441 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7621 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7681 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7741 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7801 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7861 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7921 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7981 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8041 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8101 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8161 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8221 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8281 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8341 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8401 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8461 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     8521 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     8581 cattaatgca gcggatccag acatgataag atacattgat gagtttggac aaaccacaac
     8641 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     8701 aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa
     8761 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     8821 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     8881 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg gatccgctgg
     8941 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     9001 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     9061 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     9121 gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg ggagtagtca
     9181 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     9241 aaaaaataac acaaaacaaa atat
//
