LOCUS       sequence-488207-        8766 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_188, derived from human fusion protein TCF3_FLI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8766)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8766)
  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_188
FEATURES             Location/Qualifiers
     source          1..8766
                     /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(2299..3009,3010..3012,3013..3015))
                     /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(2341..2362)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2690..2709
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2949..2970
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3012..3021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3097..3300)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3126..3146)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3301..3604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3861..3905)
                     /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    4090..4323
                     /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    4683..4800
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5292..5417
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5464..5721
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5723..6102
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6133..6237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6238..6306,6307..7098)
                     /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(6456..6475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7269..7857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7758..7777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8011..8028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8096..8230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8120..8139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8174..8193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8236..8431)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8250..8385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8490..8511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8526..8531,8532..8549,8550..8556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8564..8580
                     /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     8569..8591
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8588..8604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8588..8604
                     /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 attggccagg cgacttgaag tgggtcctaa
      601 aatctggtac ggatcaggtt gcggacgttg ctccgtgttc ttggaaattg tctgagcccc
      661 acccagcgga ggagatttat taagaccgct gttcatattg ttaccccacg ctccacggcg
      721 gacggagtca tagcttgggt cctcttttac tgacagacgt gaggattggt cggtgtgaga
      781 agtcgtgttg tacgccaaca gggaaccgct atatgagtct ggagggcgag acaggtctgg
      841 cagcgtacca ggttgtgatg gaagggctgc gtggttgtgc ataagagagg tactgcctgc
      901 cagcccatct tccggatgac tgccaccaac cagtccggcg tgacgaccac ccagtgacat
      961 aggacctgta aaacctgaag ccagagctcc gtgccctgga agtaaggtgt gcatgtcacc
     1021 agctgtgccc accgcatgac tacgtaagac atggatggcc tcgtctaagt ggtcttcgat
     1081 ctttgactgc aatccatgaa ggccaccatc ataagacggg gacagagccc ctggggcacc
     1141 cgcacgtggc cactggctcg taccagccaa accttgagga ctacctactg gtgtggaagg
     1201 actagacgag aaattattgg aagaatggtc cggtgaataa attgacgcta atgccttacc
     1261 aagggcgtct cctgaactac cagcagtcgt accacgcgat ccaagcaaag agtctgctcc
     1321 actcacaggg ggcgtgtgcg aagaaacgcc accataggtc gcgccagggg cgctagaaaa
     1381 tgatgaagcg ctaggtaaac ctccgtttac ttctgcccca tgcaattggt agcccatgcg
     1441 ctcgtgctgg tgtaacccac cgaacgtgga actggaacct gacgagccta ctgggccgga
     1501 gccgggtggc aatggcaagg gcgaagaacc acccccaagc attggtccaa accctgcctg
     1561 accaggagga gaccaaagtt ctgcagaagg atgcagcgat ccatccgcaa cgtaaaacgg
     1621 cgcgggatat gtagacgagg gagttttagc cgaagggtac gcagttgcat cgcgaccgta
     1681 gtcttcgcca gacgacggcg ggtatacact actgggcagt cctggcggca ctttacgtac
     1741 ctttttgggt tgggtgtcta atgaaccatc agccgcgcgg cgacgagagg agcccgagta
     1801 gcttgggtag tattgtgatg tgcccttcat tccggacgga gataatggcc ccggagagtt
     1861 caaggctaac tctcctgaca ggaatcccgc ctgagttaac cctcccaccc ctgcatcacg
     1921 gccaaaagaa gcgtaggcac cacgttcacc ggatttccct cccaagcctg gtcccaagaa
     1981 cgtggagctt gacagactcg aatgcgactc ggtgaaatga gtgccttccg aaaacgtgcg
     2041 tgaagggtcg aaactgcttg aactctggtc gccgctaccc caactgcccg aggacgggcg
     2101 gtcctcaaga cccgatccgc caaactgagc cccagccaag gacgcggggc gacctttacc
     2161 atttgtaacc ggcagaggaa acatcatcga aaaatccagc agatctgaca gctctttgtc
     2221 cgtgccgacg ggcgccatgc gttgaggctg gttcatccgg gaagcggccg cacttccccc
     2281 ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     2341 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     2401 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     2461 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     2521 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     2581 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     2641 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     2701 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     2761 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2821 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2881 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     2941 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     3001 gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag
     3061 aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca
     3121 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     3181 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     3241 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     3301 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     3361 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     3421 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3481 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3541 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3601 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3661 actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac
     3721 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     3781 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     3841 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     3901 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     3961 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     4021 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     4081 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     4141 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     4201 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     4261 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     4321 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     4381 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     4441 ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt
     4501 ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct
     4561 tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct
     4621 ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc
     4681 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     4741 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     4801 ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc
     4861 tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg actttgggga
     4921 ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg
     4981 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     5041 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     5101 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     5161 ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     5221 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     5281 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     5341 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     5401 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     5461 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     5521 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     5581 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     5641 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     5701 gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc
     5761 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     5821 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     5881 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     5941 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     6001 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     6061 ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt
     6121 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6181 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6241 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6301 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6361 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6421 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6481 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6541 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6601 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6661 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6721 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6781 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6841 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6901 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6961 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7021 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7081 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7141 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7201 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7261 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7321 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7381 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7441 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7501 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7561 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7621 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7681 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7741 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7801 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7861 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7921 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7981 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     8041 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     8101 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     8161 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     8221 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     8281 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     8341 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     8401 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     8461 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8521 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8581 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     8641 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     8701 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     8761 aaatat
//
