LOCUS       sequence-490714-        7275 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_043, derived from human fusion protein CTNNB1_ULK4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7275)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_043
FEATURES             Location/Qualifiers
     source          1..7275
                     /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(685..720)
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     CDS             complement(join(808..1518,1519..1521,1522..1524))
                     /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(850..871)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1199..1218
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1458..1479
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1521..1530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1606..1809)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1635..1655)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1810..2113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2370..2414)
                     /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    2599..2832
                     /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    3192..3309
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3801..3926
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3973..4230
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4232..4611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4642..4746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4747..4815,4816..5607)
                     /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(4965..4984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5778..6366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6267..6286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6520..6537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6605..6739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6629..6648)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6683..6702
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6745..6940)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6759..6894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6999..7020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7035..7040,7041..7058,7059..7065)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7073..7089
                     /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     7078..7100
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7097..7113
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7097..7113
                     /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 ttcataaagc acctgcgagg tgtctacgtc
      601 ctcctcttct ggatttcctt taccgctcag cgaaggggca gtcgtggttg ccccactatg
      661 gatgccacta tctaaataag attgttgctg ccaatggctc acagcagctt tgcggtccgg
      721 ttccatcgcc atatccaatt ccataaggtc tgcttgtgtc gccatccggg aagcggccgc
      781 acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga gagtgatccc
      841 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
      901 cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
      961 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     1021 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     1081 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1141 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1201 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     1261 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     1321 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     1381 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     1441 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1501 cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc gcggtaccgt
     1561 cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc tgcttatata
     1621 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     1681 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     1741 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     1801 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     1861 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     1921 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     1981 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2041 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     2101 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     2161 ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg
     2221 gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga aagtacagca
     2281 aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt tatataccac
     2341 agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa ttccaataat
     2401 tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg tattaagctt
     2461 gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
     2521 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
     2581 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
     2641 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
     2701 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
     2761 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
     2821 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
     2881 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
     2941 caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat gttctaatcc
     3001 tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata atccctaatg
     3061 atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat tactactgcc
     3121 ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct gctgtccctg
     3181 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     3241 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     3301 cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg ttcagcctga
     3361 tctcttacct gtcctataat tttctttaat tctttattca tagattctac tactccttga
     3421 ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg tgcttttttc
     3481 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     3541 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     3601 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     3661 tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     3721 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     3781 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     3841 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     3901 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     3961 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     4021 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     4081 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     4141 accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc
     4201 cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact aatacgtaga
     4261 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4321 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     4381 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     4441 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     4501 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     4561 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt cgacccaggt
     4621 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4681 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4741 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4801 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4861 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4921 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4981 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5041 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5101 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5161 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5221 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5281 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5341 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5401 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5461 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5521 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5581 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5641 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5701 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6421 cacatgttct ttcctgcgtt atcccctgct tctgtggata accgtattac cgcctttgag
     6481 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6541 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6601 agcggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     6661 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     6721 aagctgcaat aaacaagtta gctttttgca aaagcctagg cctccaaaaa agcctcctca
     6781 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     6841 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     6901 agttaggggc gggactatgg ttgctgacta attgagatgc ggatccgctg gcacgacagg
     6961 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7021 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7081 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgggctgcag
     7141 gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc aggagaggag
     7201 gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag aaaaaaataa
     7261 cacaaaacaa aatat
//
