LOCUS       sequence-488195-        7860 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_175, derived from human fusion protein SAFB2_SAFB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7860)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7860)
  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_175
FEATURES             Location/Qualifiers
     source          1..7860
                     /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(1393..2103,2104..2106,2107..2109))
                     /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(1435..1456)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1784..1803
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2043..2064
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2106..2115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2191..2394)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2220..2240)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2395..2698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2955..2999)
                     /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    3184..3417
                     /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    3777..3894
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4386..4511
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4558..4815
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4817..5196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5227..5331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5332..5400,5401..6192)
                     /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(5550..5569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6363..6951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6852..6871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7105..7122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7190..7324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7214..7233)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7268..7287
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7330..7525)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7344..7479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7584..7605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7620..7625,7626..7643,7644..7650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7658..7674
                     /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     7663..7685
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7682..7698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7682..7698
                     /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 ataacgacgc gtgaaacggg catcactcgg
      601 acgactgcca cgtgattgtc ctccaaatcc cccttgcatc ccgccatgcg ggatgggatg
      661 gccgcgagag gctcctccgg gcgcgaaaga cccgcgtcca gacattcctc cacggttcat
      721 catgtgccca ggcccagaat gtccagacat tgagcgctcg cccccttgcc aacgcttgtg
      781 atcacggtcc atcatccctc cgtctgctgt accctgccaa ctgcgatcat cctcgcgacg
      841 accgtggtct ccccagtcgc ggcgtccacg agggggagga ggaagaccgc gcccttcgga
      901 catacgcttg tcactgccgt agccacccca accatcacgc gaatcgcggc cgtgacgttc
      961 ggggccacca tgacgctctg ggtagtgttg tccttcgcgc tcccccatca tggaacgaga
     1021 gccctcgcgg cggtccacgg agtgatctgg gtaacgaccg cggtcgcggt ggtcgaaatc
     1081 gtgaaaacgg tcttggcgat taaaatccga gtggtaacgt tcgtctaacg cggcacgttt
     1141 agcttccggc caataggcgt cgtcgcgacg gtccaagtca tacggacggc gaactgctgg
     1201 gcgacgttcc tgttcgtagc gcagttcttg ttgacggcgt aactcttcac gttcacgatg
     1261 gatacgttct tgctcacgac gacgttcgtg ttccacatgc atgcgttcac gctccaggcg
     1321 ttcgcgttcc atgcgttcac gctctaaacg ccgggaagcg gccgcacttc ccccggagcc
     1381 tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     1441 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt
     1501 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1561 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     1621 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     1681 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     1741 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     1801 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     1861 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     1921 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     1981 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2041 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2101 gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagaattcg
     2161 gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct cccaccgtac
     2221 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     2281 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     2341 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     2401 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     2461 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     2521 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     2581 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     2641 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     2701 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagc
     2761 acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg aaacgataat
     2821 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     2881 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     2941 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     3001 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     3061 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     3121 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     3181 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     3241 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     3301 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     3361 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     3421 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     3481 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     3541 tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc ctgtctactt
     3601 gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt tgcttttctt
     3661 cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc acctttccag
     3721 aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa
     3781 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
     3841 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
     3901 atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct tacctgtcct
     3961 ataattttct ttaattcttt attcatagat tctactactc cttgactttg gggattgtag
     4021 ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact tttgttttgc
     4081 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
     4141 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
     4201 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagta
     4261 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
     4321 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
     4381 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
     4441 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
     4501 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
     4561 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     4621 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
     4681 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
     4741 tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc cccagtcccg
     4801 cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     4861 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     4921 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     4981 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     5041 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     5101 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     5161 ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga
     5221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6481 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7021 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7081 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcgg atccagacat
     7201 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     7261 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     7321 agttagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7381 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     7441 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     7501 tatggttgct gactaattga gatgcggatc cgctggcacg acaggtttcc cgactggaaa
     7561 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7621 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7681 acaggaaaca gctatgacca tgattacgcc aagcttgggc tgcaggtcga gggatctcca
     7741 taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag
     7801 taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat
//
