LOCUS       sequence-487981-        8181 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_072, derived from human fusion protein HEY1_NCOA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8181)
  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_072
FEATURES             Location/Qualifiers
     source          1..8181
                     /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(1714..2424,2425..2427,2428..2430))
                     /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(1756..1777)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2105..2124
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2364..2385
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2427..2436
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2512..2715)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2541..2561)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2716..3019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3276..3320)
                     /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    3505..3738
                     /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    4098..4215
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4707..4832
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4879..5136
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5138..5517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5548..5652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5653..5721,5722..6513)
                     /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(5871..5890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6684..7272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7173..7192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7426..7443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7511..7645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7535..7554)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7589..7608
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7651..7846)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7665..7800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7905..7926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7941..7946,7947..7964,7965..7971)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7979..7995
                     /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     7984..8006
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8003..8019
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8003..8019
                     /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 gcaatatttc cgtgttgtgt ctccctcctg
      601 cttaatcata tccattccag gcagctggtt tgggaacagg ttgcctcccc tcagagcagg
      661 atcattaacc tgctcgggac ccatggagga cagccctgac gtaggcacgg aggtcactga
      721 ggtcatgctg atctgtcctg tcatctggtt catgctgctc atgccacctg tgttggtcgc
      781 catggacaca ttgatgttca tgttgttact gtacatgctg gtgtttgctt gctgcccaaa
      841 gtgtggtggg gactgctggg aaaacatgct gtttccgccc atgttcccct gcgcccatcc
      901 atttatgtcg gagggggcct gataggctgg gttggcctga gactgttgca tcatgggact
      961 ctgtgtatgt gccattcggg gtgacataag tgggctctgg ggagtcgtag ccccagtaaa
     1021 gcctggatca ggttgctgac ttattccgta gtttggagga aatggaaact gctgtgcatt
     1081 tgcctgggga atccgagggt tgctcatagt tgctgggata ccactaggag ccaccatgct
     1141 tggtgtcata ttcaaccctt gtcctctcat catcaaagtt cgttgctgaa cttgctgttg
     1201 ctgatgcatt tgtctctgtc gaagatgctg gttcaggatt tccctctgtc tctgggccag
     1261 catctgtgca ttaataggtg cctgtgttgg tactccaggc ctcagagtca agttcacatt
     1321 ggaaacattg ctgatttgat tcataagtgg ctggcgattc tgctgtgctt ggaggcgatg
     1381 ctgaagttga agtcttagtt gatttggctg gttctgcact aggcccgtgg gcctgaggcc
     1441 cggtctgggc tgcatacgga gtgtggcata actaggccgc tgtcccatgg tgtgaaagtt
     1501 tggatcttgc atgggagaat agctaccctg ggccatttgt gcctgagatg catactgctg
     1561 tgggaaaacg ggcgccttct gctccagcat gatgttggaa tcctgacttg agaactgttc
     1621 tggatctact gcttggctct ggctgaccag ttcgggtatt cctaaggctc tccgggaagc
     1681 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1741 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1801 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1861 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1921 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1981 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     2041 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     2101 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     2161 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     2221 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     2281 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     2341 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     2401 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     2461 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     2521 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2581 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2641 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2701 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2761 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2821 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2881 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2941 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3001 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3061 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     3121 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     3181 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     3241 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     3301 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     3361 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     3421 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     3481 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     3541 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3601 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3661 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3721 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3781 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3841 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3901 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3961 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     4021 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     4081 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     4141 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     4201 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     4261 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     4321 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     4381 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     4441 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     4501 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     4561 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     4621 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     4681 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4741 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4801 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4861 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4921 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4981 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     5041 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     5101 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     5161 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     5221 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     5281 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     5341 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     5401 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     5461 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     5521 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5581 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5641 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5701 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5761 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5821 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5881 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5941 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6001 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6061 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6121 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6181 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6241 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6301 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6361 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6421 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6481 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6541 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6601 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6661 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6721 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6781 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6841 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6901 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6961 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7021 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7081 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7141 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7201 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7261 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7321 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7381 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7441 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7501 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7561 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7621 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     7681 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7741 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7801 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7861 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7921 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7981 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     8041 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     8101 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     8161 aaataacaca aaacaaaata t
//
