LOCUS       sequence-488205-        9276 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_186, derived from human fusion protein SUCO_CRTC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9276)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 9276)
  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_186
FEATURES             Location/Qualifiers
     source          1..9276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    275..392
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    752..985
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1170..1214
                     /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"
     enhancer        1471..1774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1775..1978
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1929..1949
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2054..2063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2060..2062,2063..2065,2066..2776)
                     /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(2105..2126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2366..2385)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2713..2734
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5266..5285)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5331..5386
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5385..5404)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5747..5763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5747..5763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5760..5782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5771..5787
                     /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)."
     promoter        complement(join(5795..5801,5802..5819,5820..5825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5840..5861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5920..6115
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5966..6101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6121..6255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6158..6177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6212..6231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6323..6340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6494..7082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6574..6593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7253..8044,8045..8113))
                     /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     7876..7895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8114..8218)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8249..8628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             8630..8887
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8934..9059
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
       61 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaaaaaa gcacagcaag
      121 cagcaggatc ttcagacctg gaaattccct acaatcccca aagtcaagga gtagtagaat
      181 ctatgaataa agaattaaag aaaattatag gacaggtaag agatcaggct gaacatctta
      241 agacagcagt acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
      301 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
      361 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagaaatc
      421 cactttggaa aggaccagca aagctcctct ggaaaggtga aggggcagta gtaatacaag
      481 ataatagtga cataaaagta gtgccaagaa gaaaagcaaa gatcattagg gattatggaa
      541 aacagatggc aggtgatgat tgtgtggcaa gtagacagga tgaggattag aacatggaaa
      601 agtttagtaa aacaccataa ggaggagata tgagggacaa ttggagaagt gaattatata
      661 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
      721 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
      781 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
      841 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
      901 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
      961 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1021 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1081 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1141 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1201 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1261 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1321 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     1381 ggggaccgag ctcaagcttc gaacgcgtgc tagttattaa tagtaatcaa ttacggggtc
     1441 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1501 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1561 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1621 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1681 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1741 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1801 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1861 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1921 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     1981 tttagtgaac cgggctcgtt tagtgaaccg aattctgcag tcgacggtac cgcgggcccg
     2041 ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2101 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2161 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2221 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2281 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2341 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2401 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2461 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2521 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2581 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2641 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     2701 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2761 ggacgagctg tacaagggcg gaagcggagg ctccggggga agtgcggccg cttcccggga
     2821 cgtacaattc caaaaaaaaa acacagagtc aaaaaagtta agcccacctg ttgttgagac
     2881 cttaccaaca gttgaccttc atgaggaaag cagtaatgcg gtcgtagatt cagaaactgt
     2941 tgagaatatt agctcctcaa gtacttcaga aattaccccc atttcgaaac ttgatgaaat
     3001 tgagaaaagt gggacaattc caatcgcgaa gccctcggag actgaacaat cagagacgga
     3061 ctgcgacgtc ggcgaggccc tggatgccag tgccccgatt gaacaaccta gctttgtttc
     3121 tccacccgac tctctggttg gccagcacat tgagaatgta tcaagctccc acggtaaagg
     3181 taaaatcacg aagtcggagt tcgagtctaa agtatcggcc tctgagcagg gcgggggtga
     3241 tccgaagtca gctttgaacg cgtccgataa tttgaagaat gaattgcagg cccaaaaact
     3301 gcgcttagcc tacactcgta gtagccatta cgggggctca ctgccgaacg tgaaccaaat
     3361 tgggagcgga ctggcagaat ttcagagccc acttcattca ccccttgact catctcgcag
     3421 cacacgccat catgggttag ttgagcgcgt tcagcgcgat cctcgccgca tggttagccc
     3481 actgcgccgc tacactcgtc acatcgattc gtccccttat tctccggctt acctgagccc
     3541 gccacccgaa tctagttggc gccgcacgat ggcttgggga aattttccag ctgagaaggg
     3601 tcaattgttc cgtttaccct ctgccctgaa tcgtacttcg agcgactcgg cactgcatac
     3661 ttcagtcatg aacccttccc cacaagacac gtacccaggt ccaaccccgc cttctattct
     3721 tccgtctcgt cgtggcggta tcttggacgg ggaaatggac ccgaaagtgc ctgcgattga
     3781 agaaaacttg ttggacgaca aacaccttct taagccctgg gatgccaaga agttatcgtc
     3841 ctcgtcctct cgtcctcgct catgcgaggt gccaggtatt aacatctttc cgtcgccgga
     3901 ccagccggct aatgtaccgg tgctgccccc tgcaatgaat acgggcggct cgcttccgga
     3961 cctgaccaat cttcatttcc caccaccatt accaacgcct ttggacccgg aagagaccgc
     4021 ctacccttcg ctgtcaggtg gtaattccac gtctaacctt acccacacaa tgacccacct
     4081 gggtatttca cgcgggatgg gactgggacc cggctatgac gctcccggac ttcactcacc
     4141 cctgtcccac cctagtttac agtcgagcct tagcaaccct aaccttcagg cttctttatc
     4201 atcccctcag ccgcagttac aaggctcaca cagccatcca tctcttcctg catcgtcctt
     4261 agcacgccat gtacttccca ccacttcctt agggcatcct tccctttccg caccagctct
     4321 gtcttcatct tcgtcttctt catcaacgag ttctccagta cttggagccc cttcatatcc
     4381 tgcgagcact ccgggggcgt ccccacacca ccgtcgcgtt ccattgtcgc cacttagcct
     4441 tttggcaggc cctgcagatg cgcgtcgctc acagcagcag ttacccaagc agttctcgcc
     4501 aacaatgagt cctactttgt cgagcattac gcaaggagtt cctcttgaca catccaagtt
     4561 gtccacggat cagcgcttgc ccccctaccc ctattcctca ccatcccttg tattgccaac
     4621 acaaccgcat acaccaaagt cccttcagca gcccggtctg ccatctcaat cgtgctccgt
     4681 gcaatcatca ggtggtcaac cgcccggtcg ccaatcgcat tacggaacac cctacccccc
     4741 aggaccctca ggtcacgggc aacaatccta tcaccgtcca atgagcgatt tcaatttagg
     4801 gaatctggaa cagttcagta tggaaagccc ctctgcatca ttagttcttg atccccctgg
     4861 tttctcggag ggcccagggt tcttgggggg cgaaggcccg atggggggac ctcaggaccc
     4921 tcacactttc aaccaccaaa acttaaccca ttgctctcgt catgggtcag gccctaatat
     4981 cattttgacg ggggacagta gtcctggttt ctcatgaacc ggttctagat aagattacgg
     5041 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5101 aggggggact ggaagggcta attcactccc aaagaagaca agatctgctt tttgcctgta
     5161 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5221 cactgcttaa gcctcaataa agcttcagct gctcgagcta gcagatcttt ttccctctgc
     5281 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     5341 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     5401 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     5461 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     5521 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     5581 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     5641 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     5701 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     5761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5881 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     5941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6241 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8221 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa
     8281 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     8341 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     8401 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     8461 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     8521 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     8581 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg aggcgtggcc
     8641 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     8701 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     8761 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     8821 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     8881 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     8941 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     9001 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     9061 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     9121 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     9181 ttaatactgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     9241 catcccttca gacaggatca gaagaactta gatcat
//
