LOCUS       sequence-488208-        8613 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_189, derived from human fusion protein TCF3_HLF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8613)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8613)
  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_189
FEATURES             Location/Qualifiers
     source          1..8613
                     /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(2146..2856,2857..2859,2860..2862))
                     /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(2188..2209)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2537..2556
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2796..2817
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2859..2868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2944..3147)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2973..2993)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3148..3451
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3708..3752)
                     /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    3937..4170
                     /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    4530..4647
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5139..5264
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5311..5568
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5570..5949
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5980..6084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6085..6153,6154..6945)
                     /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(6303..6322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7116..7704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7605..7624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7858..7875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7943..8077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7967..7986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8021..8040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8083..8278)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8097..8232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8337..8358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8373..8378,8379..8396,8397..8403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8411..8427
                     /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     8416..8438
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8435..8451
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8435..8451
                     /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 ctggttctcc ttcaggcgac gcgcatcacg
      601 gctgcgtttt gccgccatgt tgttcttgcg gcggcgcgcc cagtacttat catcaaccgt
      661 cggcgtcgga agtcccaaac atgttaatga gggggctggt tgagagggca aagccgcatg
      721 gttgtacata agtgatgtac tgccagccaa tccatcttcg gggtgagacc ccccaaccag
      781 gccggcatgg cgccctccca gggacattgg accagtgaac cctgatgcca gcgctccgtg
      841 ccctggtaac aatgtgtgca tgtctccggc cgtgccgaca gcatgactac gaagtacatg
      901 gatggcctca tccagatgat cttcgatttt gctctgaagc ccatgcagcc ccccatcgta
      961 agagggcgat aaagctcccg gagcgcccgc acgcggccat tgggatgtcc cggccaaccc
     1021 ctgtggagac cctaccggag tacttggtga agatgagaag ttattagatg agtggtctgg
     1081 tgaatagata ctagcaagtg cctttccaag tgcgtcaccg ctggaacctg cagtggtacc
     1141 gcgtgaaccc agcaaagaat cagcacctga gacagggggt gtatgtgacg aaactccccc
     1201 gtatgtagcg cctggggcgc tgctaaacga gctcgcggat ggtaaacccc cgtttacctc
     1261 agcgccgtgt aattgatacc ccatgcgttc gtgttggtga agccccccga aggtcgaact
     1321 cgacccggag ctcccgactg gcccgctacc tgggggcaga ggaagaggag aagagccccc
     1381 gcctaacatc gggccaaatc cggcctgtcc tggagggctc cacagctctg ctgaaggatg
     1441 caggcttcca tccgctacgt agaaaggggc agggtaagtt gaggaaggag ttttggccga
     1501 tgggtaagct gttgcgtcac ggccataatc ctcgcctgaa gaaggggggt agacactact
     1561 cggtaagcca ggaggcactt tacgcacttt cttaggttga gtgtctaagg aaccatcagc
     1621 tgcgcgacgg cgagaggacc cactatagct agggtagtat tgggacgtcc ccttcattcc
     1681 tgaagggcta agtggtccgg ggctgttcaa agcaagttca ccactcagga accccgcttg
     1741 tgtcaaacca cctactcccg catcacggcc gaaagaggca tacgcgccgc gctcaccgct
     1801 ctttccgccc aaacccggac caagaaaagt actagaactc aaggaactgt ggctttcggt
     1861 gaagtgagta ccttctgaaa acgtgcggct tggatcgaag gatgaagagc tttgatcacc
     1921 agacccccat gatccgctgc ttggacggtc ttcaaggccc gaccccccaa actgggcccc
     1981 tgcaagagag gccgggcgcc ccttgccatt ggttacgggc aaagggaaca tcatagagaa
     2041 atcaagtaaa tcactaagct ccttgtcagt accaactggg gccatgcgct gaggttggtt
     2101 catccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
     2161 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     2221 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     2281 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     2341 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     2401 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     2461 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     2521 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     2581 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     2641 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     2701 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     2761 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     2821 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     2881 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     2941 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     3001 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     3061 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     3121 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     3181 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     3241 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     3301 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     3361 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     3421 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     3481 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     3541 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     3601 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     3661 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     3721 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     3781 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     3841 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     3901 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     3961 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     4021 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     4081 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     4141 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     4201 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     4261 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     4321 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     4381 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     4441 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     4501 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     4561 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     4621 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     4681 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     4741 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     4801 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     4861 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     4921 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     4981 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     5041 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     5101 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     5161 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     5221 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     5281 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     5341 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     5401 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     5461 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     5521 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     5581 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     5641 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     5701 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     5761 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     5821 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     5881 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     5941 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6001 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6061 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6121 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6181 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6241 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6301 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6361 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6421 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6481 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6541 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6601 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6661 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6721 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6781 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6841 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6901 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6961 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7021 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7081 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7141 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7201 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7261 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7321 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7381 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7441 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7501 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7561 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7621 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7681 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7741 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7801 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7861 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7921 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     7981 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     8041 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     8101 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     8161 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     8221 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     8281 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8341 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8401 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8461 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     8521 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     8581 tgttaaagaa aaaaataaca caaaacaaaa tat
//
