LOCUS       sequence-487949-        7461 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_035, derived from human fusion protein CLK1_PPIL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7461)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7461)
  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_035
FEATURES             Location/Qualifiers
     source          1..7461
                     /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(994..1704,1705..1707,1708..1710))
                     /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(1036..1057)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1385..1404
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1644..1665
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1707..1716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1792..1995)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1821..1841)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1996..2299
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2556..2600)
                     /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    2785..3018
                     /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    3378..3495
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3987..4112
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4159..4416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4418..4797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4828..4932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4933..5001,5002..5793)
                     /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(5151..5170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5964..6552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6453..6472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6706..6723
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6791..6925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6815..6834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6869..6888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6931..7126)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6945..7080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7185..7206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7221..7226,7227..7244,7245..7251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7259..7275
                     /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     7264..7286
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7283..7299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7283..7299
                     /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 cagatgcccc tcttcatcat cttcaacgct
      601 acgagtgcgt ttgcgacggt gagacttacc atgcgaacgg cggtggcttg tgctatgatg
      661 gatgcggtgc ttagatttgt acgatgagcg ccctgagcgg ccagacgact tagatgaatg
      721 attctgatag cgactttcat ggtcacgttg gcgatgtcct ggttcgcacc cctgtgtgta
      781 gtcgttgcga tactcatcaa tatagcgacg agaatgatag tccttttcat tgatcgaacg
      841 agactccaaa taatgagaat cgcacatctt gctgtgatta tatttacaac gcttgttctc
      901 ttgcgccgaa ctatgagaac gtttacgacg tttgtgggaa gagctggagc gccgggaagc
      961 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1021 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1081 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1141 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     1201 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     1261 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     1321 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1381 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1441 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     1501 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1561 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1621 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1681 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     1741 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     1801 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     1861 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     1921 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     1981 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2041 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2101 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2161 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2221 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2281 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2341 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     2401 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     2461 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     2521 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     2581 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     2641 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     2701 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     2761 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     2821 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     2881 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     2941 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3001 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3061 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3121 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     3181 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     3241 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     3301 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     3361 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     3421 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     3481 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     3541 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     3601 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     3661 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     3721 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     3781 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     3841 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     3901 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     3961 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4021 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4081 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4141 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     4201 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     4261 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     4321 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     4381 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     4441 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4501 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4561 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4621 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4681 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     4741 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     4801 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4861 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4921 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4981 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5041 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5101 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5161 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5221 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5281 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5341 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5401 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5461 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5521 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5581 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5641 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5701 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5761 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5821 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5881 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5941 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6001 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6061 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6121 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6181 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6241 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6301 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6361 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6421 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6481 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6541 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6601 tttgctcaca tgttctttcc tgcgttatcc cctgcttctg tggataaccg tattaccgcc
     6661 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6721 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6781 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6841 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6901 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     6961 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7021 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7081 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7141 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7201 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7261 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     7321 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     7381 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     7441 aaataacaca aaacaaaata t
//
