LOCUS       sequence-487969-        7581 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_058, derived from human fusion protein EP400_SFSWAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7581)
  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_058
FEATURES             Location/Qualifiers
     source          1..7581
                     /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(1114..1824,1825..1827,1828..1830))
                     /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(1156..1177)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1505..1524
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1764..1785
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1827..1836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1912..2115)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1941..1961)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2116..2419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2676..2720)
                     /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    2905..3138
                     /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    3498..3615
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4107..4232
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4279..4536
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4538..4917
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4948..5052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5053..5121,5122..5913)
                     /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(5271..5290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6084..6672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6573..6592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6826..6843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6911..7045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6935..6954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6989..7008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7051..7246)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7065..7200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7305..7326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7341..7346,7347..7364,7365..7371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7379..7395
                     /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     7384..7406
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7403..7419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7403..7419
                     /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 tgtcggggat gagctgcaaa gaccgaggcc
      601 cggacccggc gcaccggcac gaacgttttg cagtgcttgg cccggacccg gtgacggctg
      661 cgtcgggctt tgggtctgca cttgttggct caacggcgag gtcacctgaa tataggacgg
      721 ggagccgtgc tcgaaacggc gaggctgcgc gctgaactgg aagcccggag aagtcgggct
      781 cggcaacggc agcggggcca acgtaatctg ctggttgcct ccaacaaccg ggcccacact
      841 ctgcagggta atattcacgt tctggccggt agccgggcta cgattcatca gttgttgaat
      901 ctggtagctc gggctctgcg gcgcagacgg gctagcagac ggtgcaaatg gagccgccgg
      961 cgatggcggc ggattcgggt gggccggctg ctcttcacct tcgctgcccg gacatgcacg
     1021 agaacgttgg agctggtgct gtacattttg gggtcctgtc ccgtgatgca tccgggaagc
     1081 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1141 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1201 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1261 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1321 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1381 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1441 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1501 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1561 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     1621 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1681 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1741 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1801 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     1861 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     1921 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     1981 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2041 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2101 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2161 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2221 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2281 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2341 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2401 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2461 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     2521 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     2581 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     2641 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     2701 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     2761 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     2821 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     2881 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     2941 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3001 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3061 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3121 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3181 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3241 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3301 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3361 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     3421 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     3481 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     3541 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     3601 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     3661 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     3721 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     3781 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     3841 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     3901 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     3961 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     4021 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     4081 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4141 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4201 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4261 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4321 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4381 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     4441 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     4501 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     4561 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4621 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4681 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4741 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4801 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4861 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     4921 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4981 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5041 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5101 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5161 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5221 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5281 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5341 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5401 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5461 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5521 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5581 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5641 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5701 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5761 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5821 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5881 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5941 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6001 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6061 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6721 tttgctcaca tgttctttcc tgcgttatcc cctgcttctg tggataaccg tattaccgcc
     6781 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6841 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6901 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7021 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     7081 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7141 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7201 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7261 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7321 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7381 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     7441 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     7501 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7561 aaataacaca aaacaaaata t
//
