LOCUS       sequence-488183-        7632 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_163, derived from human fusion protein RBM28_PRRT4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7632)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7632)
  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_163
FEATURES             Location/Qualifiers
     source          1..7632
                     /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(1165..1875,1876..1878,1879..1881))
                     /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(1207..1228)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1556..1575
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1815..1836
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1878..1887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1963..2166)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1992..2012)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2167..2470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2727..2771)
                     /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    2956..3189
                     /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    3549..3666
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4158..4283
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4330..4587
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4589..4968
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5104..5172,5173..5964)
                     /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(5322..5341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6877..6894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6962..7096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6986..7005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7040..7059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7102..7297)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7116..7251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7356..7377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7392..7397,7398..7415,7416..7422)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7430..7446
                     /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     7435..7457
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7454..7470
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7454..7470
                     /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 ctccaacgta aggaaaaaca atgggcgggg
      601 agcctcggga ggttgccctt cgcgttcagg caattcgcct accgagcagg agcctgggcc
      661 acattcgggt tgtcctgagg gatttcctaa agaggccggg ggtgataagt cgtcaggccc
      721 gctgataggc accgtagggt ctttgtcctc ttgttcttcg gttgacgtat ctgactgagc
      781 aagttcttct ccgtcatcga tgctatcgga ttcttccaag tctgaatctt cttctgagtg
      841 atcagacgac ttcgcagggg cgggacgctt cactgcgcgc ttttgaatct gtaccggctt
      901 tgtcactttg gattcgatat tctcttcctc ttcatcctca tcgtcaaata caccatcctc
      961 ctcatcatca tcgtcatcat catcatcatt ttcttcttct tccatatctt cttcttcacg
     1021 gcctttcttt ttcacgcttt cctgatgctt ggattcatga gatttctctt caccaatagc
     1081 tgatacggat tgagtatcct tatacttatc tttagctact gcccgggaag cggccgcact
     1141 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
     1201 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1261 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1321 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1381 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1441 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1501 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1561 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1621 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1681 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1741 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1801 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1861 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     1921 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     1981 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2041 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2101 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2161 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2221 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2281 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2341 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2401 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2461 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2521 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     2581 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     2641 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     2701 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     2761 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     2821 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     2881 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     2941 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     3001 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     3061 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     3121 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     3181 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     3241 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     3301 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3361 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3421 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3481 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3541 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     3601 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     3661 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     3721 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     3781 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     3841 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     3901 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     3961 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     4021 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     4081 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     4141 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     4201 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     4261 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     4321 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4381 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4441 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4501 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4561 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     4621 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4681 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4741 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4801 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4861 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4921 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5161 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5341 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5641 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6241 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6781 atgttctttc ctgcgttatc ccctgcttct gtggataacc gtattaccgc ctttgagtga
     6841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6901 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6961 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     7021 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7081 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     7141 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     7201 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     7261 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     7321 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7381 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7441 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7501 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7561 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     7621 aaaacaaaat at
//
