LOCUS       sequence-488222-        8163 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_205, derived from human fusion protein UBXN4_R3HDM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8163)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8163)
  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_205
FEATURES             Location/Qualifiers
     source          1..8163
                     /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(1696..2406,2407..2409,2410..2412))
                     /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(1738..1759)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2087..2106
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2346..2367
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2409..2418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2494..2697)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2523..2543)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2698..3001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3258..3302)
                     /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    3487..3720
                     /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    4080..4197
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4689..4814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4861..5118
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5120..5499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5530..5634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5635..5703,5704..6495)
                     /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(5853..5872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6666..7254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7155..7174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7408..7425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7493..7627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7517..7536)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7571..7590
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7633..7828)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7647..7782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7887..7908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7923..7928,7929..7946,7947..7953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7961..7977
                     /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     7966..7988
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7985..8001
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7985..8001
                     /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 atatttattt tgtttccact gcgggggtga
      601 atgcgcacag gattgcgggt tgttgggtac gcttaactgc atcattacca ttccgccccc
      661 aggaggaggt ggcggcccag ccgtgcactg gtgaccctga agctgctgag aggagcatgg
      721 gctggtggta cggggaaact gtacttgctc gctgccaggg tgctgcaaat aacctaccga
      781 agaactcaag ttattttgtt ggccaggggg gatgacagag tagtataccg gcatgcctgt
      841 tgttggcatg gagccttggt tggactgatt gggaaacatt acaggttgct gatacgtttg
      901 atggggaata ccttgcgacc cttggggtaa cgacacttgg gccgcaaact gacgcgcttc
      961 ctcaagcggt gcgtcgctgg gaaactgatt cgtaaaagaa gagccatctg gtaagcgaaa
     1021 ttggatacgc gcaaccgttg agcgttcacg agcataagat tcacgtttta cttccatttc
     1081 tgcttgctta gccaacaatg ccgctgcttt tgcggcctcg acttcttctt tggtctttgc
     1141 aaagcgagca gcacgctcag cacggtccaa ggcaatttgc tgcttaatgc gctcgcgcgc
     1201 cgcacggtct tcggcctttt cgcgattgcg ctcttctaac atacgtttag taagctcctc
     1261 ctcttgcttg cgcttatagt ctaacatttc cttgcccgtt ttgcgacgct cgatctcctt
     1321 cttaatttca cgctgttctt cctccttacg tttttcctcg cggcgttctt ccaacttttt
     1381 ggtaagacgt tctacgcgaa tatttaaatc ttccgcggga cgctggtcgg aacacccaga
     1441 cggttcctga gacgatgtcg cgtgtccggc ggattctcca cccgttgcgg tatctgattt
     1501 tgtatcagat gtgggtggga tttcgcaaag ctccgcatta cgtgactgtg aattttcgca
     1561 ggtgttgttc ggttcaaaac ttgcagaagg agttgacacg gatgattcac tctgagaacc
     1621 gttcgccacg gacgtctcac ttttaagcag gtgccgggaa gcggccgcac ttcccccgga
     1681 gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
     1741 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
     1801 ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     1861 ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt
     1921 caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt
     1981 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     2041 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     2101 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     2161 gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag
     2221 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag
     2281 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     2341 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     2401 cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat
     2461 tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga cctcccaccg
     2521 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     2581 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     2641 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     2701 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     2761 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     2821 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     2881 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     2941 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     3001 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     3061 agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat
     3121 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     3181 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     3241 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     3301 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     3361 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     3421 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     3481 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     3541 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     3601 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     3661 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     3721 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     3781 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     3841 atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc atcctgtcta
     3901 cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat ctttgctttt
     3961 cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc ttcacctttc
     4021 cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta ataaacccga
     4081 aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt
     4141 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg
     4201 tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc tcttacctgt
     4261 cctataattt tctttaattc tttattcata gattctacta ctccttgact ttggggattg
     4321 tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt acttttgttt
     4381 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     4441 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     4501 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     4561 gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     4621 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     4681 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     4741 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     4801 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     4861 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     4921 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     4981 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     5041 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
     5101 ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg tactgccaag
     5161 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     5221 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     5281 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     5341 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     5401 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     5461 taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg cacttttcgg
     5521 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5581 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5641 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5701 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5761 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5821 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5881 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5941 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6001 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6061 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6121 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6181 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6241 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6301 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6361 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6421 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6481 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6541 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6601 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6661 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6721 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6781 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6841 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6901 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6961 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7021 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7081 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7141 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7201 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7261 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7321 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7381 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7441 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag cggatccaga
     7501 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     7561 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     7621 acaagttagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     7681 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     7741 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     7801 gactatggtt gctgactaat tgagatgcgg atccgctggc acgacaggtt tcccgactgg
     7861 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7921 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7981 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     8041 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     8101 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     8161 tat
//
