LOCUS       sequence-488194-        7710 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_174, derived from human fusion protein SAFB2_SAFB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7710)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7710)
  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_174
FEATURES             Location/Qualifiers
     source          1..7710
                     /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(1243..1953,1954..1956,1957..1959))
                     /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(1285..1306)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1634..1653
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1893..1914
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1956..1965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2041..2244)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2070..2090)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2245..2548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2805..2849)
                     /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    3034..3267
                     /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    3627..3744
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4236..4361
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4408..4665
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4667..5046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5077..5181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5182..5250,5251..6042)
                     /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(5400..5419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6213..6801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6702..6721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6955..6972
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7040..7174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7064..7083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7118..7137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7180..7375)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7194..7329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7434..7455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7470..7475,7476..7493,7494..7500)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7508..7524
                     /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     7513..7535
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7532..7548
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7532..7548
                     /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 gcgctcctca cgctcccatt gggcctgcat
      601 acgttgctca cgctccgagc gctcacgaac acgactgtgg ctttctgaat cacgtgactt
      661 gcgcggttcc ttcaccttgt cgaaggagac tactgagcgt ttctcacgag acgcagactt
      721 acggtcttga ctttttgatg cacgctcctt cgatccacta gtcttgacac taatgacagg
      781 caccccttta gacttgtcca taactactgt gcgttcggtc ccgcggcttc cagatttagt
      841 ggcacgtgag cgttcggacg ggcctggctt ctggtcgtct tggtccttag atttttcacc
      901 gctcccgtcg tccccctttt tagcgtcgtc tttacgatcg catttgtcgt cacgttttaa
      961 gttggtcgaa cggtccgaat tactagactt ctccttctta ccgtcagagt cgcgtttatc
     1021 cgacgttttt ttgcccaccg gttcgttttt agccttttcc acagagatca ttttgccgtg
     1081 aagctcggtt ttgtgtaagt ggttgataca tttagtggct tcctcagcgg tggacattgt
     1141 cacgaagccg tagcagcgcg ctcctggcga acgagcatta gtgactactt tggcccccac
     1201 ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca gctcgtccat
     1261 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     1321 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
     1381 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1441 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     1501 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1561 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1621 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1681 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1741 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     1801 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1861 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1921 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
     1981 ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt tcactaaacg
     2041 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     2101 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     2161 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     2221 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     2281 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     2341 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     2401 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     2461 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     2521 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     2581 ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct cggtcccctc
     2641 ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac tctattcact
     2701 atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat cattatgaat
     2761 aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact tgcccattta
     2821 tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc ttcaattaag
     2881 gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca ggtcgtgtga
     2941 ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac agcagtggtg
     3001 caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt taggtatctt
     3061 tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag ttgcaacaga
     3121 tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat accagacaat
     3181 aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct
     3241 cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct ctgcaccact
     3301 cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac tttatattta
     3361 tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt actaaacttt
     3421 tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc catctgtttt
     3481 ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc actattatct
     3541 tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt ccaaagtgga
     3601 tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     3661 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     3721 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac tgctgtctta
     3781 agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt attcatagat
     3841 tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga tcctgctgct
     3901 tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     3961 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     4021 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     4081 cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag ctccctgctt
     4141 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     4201 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     4261 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     4321 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     4381 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     4441 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     4501 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     4561 ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc ttatatgcag
     4621 gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg gtaatagcga
     4681 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     4741 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     4801 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     4861 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     4921 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     4981 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     5041 aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5161 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5221 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5281 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5341 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5401 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5461 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5521 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5581 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5641 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5701 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5761 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5821 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5881 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5941 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6001 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6061 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6121 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6181 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6301 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6361 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6421 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6481 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6541 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6601 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6661 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6721 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6781 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6841 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6901 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6961 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7021 ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt ttggacaaac
     7081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     7141 atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc ctaggcctcc
     7201 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     7261 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     7321 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcggatc
     7381 cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7441 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7501 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7561 aagcttgggc tgcaggtcga gggatctcca taagagaaga gggacagcta tgactgggag
     7621 tagtcaggag aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt
     7681 taaagaaaaa aataacacaa aacaaaatat
//
