LOCUS       sequence-487961-        7995 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_048, derived from human fusion protein CUX1_NUTM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7995)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7995)
  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_048
FEATURES             Location/Qualifiers
     source          1..7995
                     /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(1528..2238,2239..2241,2242..2244))
                     /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(1570..1591)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1919..1938
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2178..2199
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2241..2250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2326..2529)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2355..2375)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2530..2833
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3090..3134)
                     /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    3319..3552
                     /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    3912..4029
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4521..4646
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4693..4950
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4952..5331
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5362..5466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5467..5535,5536..6327)
                     /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(5685..5704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6498..7086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6987..7006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7240..7257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7325..7459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7349..7368)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7403..7422
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7465..7660)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7479..7614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7719..7740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7755..7760,7761..7778,7779..7785)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7793..7809
                     /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     7798..7820
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7817..7833
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7817..7833
                     /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 agttgccaaa tcctcgatcg ttttcgggct
      601 cgggtagggt ttttgttgat atgcgcgttt cagggcttcc ttctcttccg gcgcgaggac
      661 cacacgcggc ttctttaatt gatgctgcgg ttgcgggctc gcaccctggc tatactcggt
      721 gcctacactc ggcggttcgc acggctggct gtcgctaacg ctggaatgac gtctcttcat
      781 atacgccttc ttttccatcc gtttcatgtc cattaacttc tccacgttat tcggatcgtt
      841 cagccaaagc tgcatacgca cgaacggttc acgacccttt aaggacagtt tatgccacgg
      901 cttcgggcgc gccagcaggt cggatacgct accttgagtc agacctaaaa tggtttcacc
      961 aaacaggcgc tgacccaggt tgttatcggt cagaacctct ttaacacgtt tggtaatacc
     1021 ataggtgtcc agttccgggc tcatggcgac cagctcttga attgaaagtg cactgtggcc
     1081 ggatagcggg agcggagtag tcggttggct gtcagacgcc ggcgggtcgc tcggctccgg
     1141 cggtttgctg tccttgctcg cctcaatcgg cgggcatggt tgctgcacca actcagtcag
     1201 gctcttcaca gactctgagg aactcatcgg ggattcagga gccgggctgc atgacgctga
     1261 ggttttcggg gtggattcgg aggaaacgca cccttgttgt agcgggtcct ggagagaggt
     1321 caccgagtgc aagacgggac cttgttgctg accctgaacc ggcaacacgc cttgacccaa
     1381 ttcgccgttc agccacagct gcatgcggat gaacggttca cggccttttt gggtcagctt
     1441 tgaccaaggt ttcggacggg acagcatatc cgaaactgat ccctgccggg aagcggccgc
     1501 acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga gagtgatccc
     1561 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     1621 cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     1681 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     1741 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     1801 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1861 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1921 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     1981 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     2041 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     2101 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     2161 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     2221 cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc gcggtaccgt
     2281 cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc tgcttatata
     2341 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2401 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2461 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2521 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2581 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2641 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     2701 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2761 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     2821 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     2881 ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg
     2941 gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga aagtacagca
     3001 aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt tatataccac
     3061 agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa ttccaataat
     3121 tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg tattaagctt
     3181 gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
     3241 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
     3301 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
     3361 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
     3421 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
     3481 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
     3541 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
     3601 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
     3661 caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat gttctaatcc
     3721 tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata atccctaatg
     3781 atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat tactactgcc
     3841 ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct gctgtccctg
     3901 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     3961 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     4021 cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg ttcagcctga
     4081 tctcttacct gtcctataat tttctttaat tctttattca tagattctac tactccttga
     4141 ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg tgcttttttc
     4201 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     4261 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     4321 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     4381 tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     4441 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     4501 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     4561 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     4621 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     4681 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     4741 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     4801 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     4861 accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc
     4921 cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact aatacgtaga
     4981 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     5041 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     5101 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     5161 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     5221 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     5281 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt cgacccaggt
     5341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5461 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5521 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5581 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5641 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5701 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5761 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5821 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5881 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5941 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6001 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6061 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6121 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6181 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6241 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6301 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6361 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6421 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6481 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6541 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6601 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6661 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6721 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6781 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6841 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6901 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6961 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7021 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7081 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7141 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7201 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7261 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7321 agcggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     7381 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     7441 aagctgcaat aaacaagtta gctttttgca aaagcctagg cctccaaaaa agcctcctca
     7501 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     7561 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     7621 agttaggggc gggactatgg ttgctgacta attgagatgc ggatccgctg gcacgacagg
     7681 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7741 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7801 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgggctgcag
     7861 gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc aggagaggag
     7921 gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag aaaaaaataa
     7981 cacaaaacaa aatat
//
