LOCUS       sequence-488209-        8721 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_190, derived from human fusion protein TCF3_PBX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8721)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8721)
  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_190
FEATURES             Location/Qualifiers
     source          1..8721
                     /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(2254..2964,2965..2967,2968..2970))
                     /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(2296..2317)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2645..2664
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2904..2925
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2967..2976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3052..3255)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3081..3101)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3256..3559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3816..3860)
                     /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    4045..4278
                     /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    4638..4755
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5247..5372
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5419..5676
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5678..6057
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6088..6192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6193..6261,6262..7053)
                     /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(6411..6430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7224..7812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7713..7732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7966..7983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8051..8185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8075..8094)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8129..8148
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8191..8386)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8205..8340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8445..8466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8481..8486,8487..8504,8505..8511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8519..8535
                     /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     8524..8546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8543..8559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8543..8559
                     /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 cgcacggtaa tcggagtgtt caaccgagtt
      601 gtccgaaccg gcgccaccgg aggccgctgc agcagctgcc gctgctgcgc tacccccgcc
      661 cttctccggt cctgcaacac cctcggccaa caacatgttg tccagacgca tcaattgtgg
      721 gtcagtaggt tcttcctcct gagcgccgcg aatggacagg acgctgtacg aatccggcgg
      781 gcgactcaga tcgggaagag tgcccggttg gctaggcagc gctgcgtggt tatgcatcaa
      841 tgacgtcgaa cccgctaatc cgtcttccgg gtgggagccg ccgactaaac cagcgtgacg
      901 tccccccagt gacataggac ctgtgaagcc agatgccagc gcgccgtgcc caggtaacaa
      961 tgtatgcata tcgccagctg ttcctaccgc gtgcgagcga agtacgtgaa tcgcctcatc
     1021 caggtgatcc tcaatcttag actgcaatcc gtgtaagccc ccgtcgtatg aaggactcaa
     1081 agcaccaggt gcccccgcgc ggggccattg tgacgtcccg gctaaccctt gaggacttcc
     1141 aacgggtgtt gagggggatg aactgaagtt gttactagag tggtcaggtg agtaaatgct
     1201 cgccagagcc ttgcctaagg catcaccaga actccccgcg gtggtcccac gggaaccaag
     1261 cagactatcc gcaccactta ccgggggagt gtgcgacgag acccctccgt aagtggcacc
     1321 gggggcgctg gaaaaagacg atgcagaggg cagcccgccg ttgacttcag ctccatgtaa
     1381 ttgatacccc atacgctcgt gctggtgaag tcccccgaaa gtcgaagaac tgccggaact
     1441 gcctacaggc ccgctacccg gaggaagtgg caaaggactc gacccgccac ccaacatcgg
     1501 accaaatccc gcttggccag gcggcgacca aagctccgca gatgggtgta acgagccgtc
     1561 cgccacgtaa aacggggccg gataagtact tgaaggagtt ttggcagatg ggtacgcggt
     1621 cgcatcgcgt ccatagtcct ctcctgaact tgggggatag accgaactcg gtaatccggg
     1681 aggaactttg cgcacctttt tgggctgggt atcaaggctg ccatcggctg cacgacgacg
     1741 cgacgaaccg ctgtaagaag gataatattg agaggttcct ttcattccac ttggcgaaag
     1801 agggccgggg gaatttaatg ctaactcacc gctaagaaag cccgcctgtg taaggccacc
     1861 tactcccgca tcacggccaa aactggcata agccccacgt tcccccgact taccgccaag
     1921 gcccggcccc aggaaagttg agctggacaa ggatgaatgg gattcggtaa agtgggtccc
     1981 ttcagaaaaa gtgcgggatg ggtcaaagga ggaagatgac tgatccccgg aaccccagct
     2041 cccagatgag gggcgatcct ccaggccgct gcctccgaac tgggctcccg ccaatgatgc
     2101 ggggcgccct ttaccattag tcaccggcag cggaaacatc atcgagaaat ccaataagtc
     2161 tgataattct ttatccgttc ccactggggc catgcgttgg ggctggttca tccgggaagc
     2221 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     2281 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2341 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2401 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     2461 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     2521 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     2581 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     2641 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     2701 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     2761 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     2821 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     2881 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     2941 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     3001 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     3061 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     3121 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     3181 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     3241 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     3301 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     3361 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     3421 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     3481 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3541 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3601 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     3661 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     3721 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     3781 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     3841 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     3901 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     3961 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     4021 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     4081 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     4141 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     4201 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     4261 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     4321 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     4381 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     4441 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     4501 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     4561 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     4621 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     4681 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     4741 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     4801 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     4861 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     4921 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     4981 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     5041 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     5101 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     5161 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     5221 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     5281 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     5341 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     5401 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     5461 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     5521 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     5581 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     5641 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     5701 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     5761 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     5821 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     5881 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     5941 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     6001 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     6061 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6181 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6241 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6301 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6361 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6421 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6481 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6541 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6601 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6661 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6721 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6781 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6841 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6901 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6961 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7021 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7081 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7981 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8041 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     8101 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     8161 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     8221 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     8281 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     8341 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     8401 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8461 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8521 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     8581 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     8641 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     8701 aaataacaca aaacaaaata t
//
