LOCUS       sequence-488237-        7965 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_007, derived from human protein FUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7965)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7965)
  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_Vidr_007
FEATURES             Location/Qualifiers
     source          1..7965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(2996..3706,3707..3709,3710..3712))
                     /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(3038..3059)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3387..3406
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3646..3667
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3709..3718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3794..3997)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3823..3843)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3998..4301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4558..4602)
                     /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    4787..5020
                     /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    5380..5497
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5989..6114
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6161..6418
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6420..6799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6830..6934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6935..7003,7004..7795)
                     /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(7153..7172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct gctagctcga gcagctgaag ctttattgag gcttaagcag
     1861 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
     1921 agtacaggca aaaagcagat cttgtcttct ttgggagtga attagccctt ccagtccccc
     1981 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg
     2041 taccgtaatc ttatctagaa ccggttcaat tctccccaag gccctgaaca aaaatggtgt
     2101 tgttgtcact attgtcttgt tcgctgtcgt ggcgggagcc ttgatctcgt ggaccgccaa
     2161 acttattgaa accgcctcgg tcgctcccac ccattcctcc tcgacctcct cggcctcctc
     2221 cgcgacccct aggttcgtac ccgccggatg acctgttgta tcctccgcca ccaccaccac
     2281 ctcccccacc acttcctccc cgacctctgc cccctctatc ctgttgccca tagccaccac
     2341 tccccccacc tcctgattga tcttgattac cgtatccgcc gcctgaccct ccgccagaag
     2401 acatggaact ctgatcctga ccatagttgc caccgcctcc accaccacct ccgccgccac
     2461 tacttgagtt atactgattc tgttggccat acccttgagg gggattataa gactgctgtt
     2521 gcccataaga ttgttgttgg ccgccatatg atggttgctg tgaatatgaa ccagactgag
     2581 gttgcccata tgaacttgat tgggaacttg acccatagct gcctgaagtg gaactaggag
     2641 cgggttgctg cccatatcct ggataagatg actgctgacc ataactagat tgacttgact
     2701 gtgagctgcc atatccgcct gtagaaccgt atccctgtgg ggtactttgg gtgccgtacc
     2761 ccgtgttctg gctctgaccg taagatgaat agcttgattg accgtaacct gaggtatcgg
     2821 tgctctggga ataccctgag taactttgtt ggccataagg ttggctgctc tgttgactgt
     2881 agccctgacc gggctgcgtc ggatacgcac cgtagctttg cgtggcttgc tgagtatagt
     2941 cgttggacgc catccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     3001 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     3061 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     3121 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     3181 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     3241 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     3301 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     3361 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     3421 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     3481 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     3541 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     3601 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3661 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     3721 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     3781 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     3841 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     3901 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     3961 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4021 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4081 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4141 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4201 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4261 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4321 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     4381 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     4441 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     4501 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     4561 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     4621 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     4681 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     4741 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     4801 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     4861 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     4921 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     4981 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     5041 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     5101 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     5161 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     5221 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     5281 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     5341 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     5401 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     5461 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     5521 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     5581 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     5641 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     5701 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     5761 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     5821 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     5881 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     5941 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     6001 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     6061 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     6121 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     6181 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     6241 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     6301 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     6361 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     6421 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6481 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6541 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6601 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6661 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6721 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     6781 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
