LOCUS       sequence-488213-        7443 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_194, derived from human fusion protein THSD4_LRRC49.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7443)
  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_194
FEATURES             Location/Qualifiers
     source          1..7443
                     /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(976..1686,1687..1689,1690..1692))
                     /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(1018..1039)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1367..1386
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1626..1647
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1689..1698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1774..1977)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1803..1823)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1978..2281
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2538..2582)
                     /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    2767..3000
                     /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    3360..3477
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3969..4094
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4141..4398
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4400..4779
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4810..4914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4915..4983,4984..5775)
                     /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(5133..5152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5946..6534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6435..6454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6688..6705
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6773..6907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6797..6816)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6851..6870
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6913..7108)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6927..7062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7167..7188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7203..7208,7209..7226,7227..7233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7241..7257
                     /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     7246..7268
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7265..7281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7265..7281
                     /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 gccactgtca agcggacccg tctcctcggg
      601 aaacgccgat aaagttgacc cgtcgatctg gcatgggtcc tgtggactat cagaatcttt
      661 gcgatcctca ttagtggcaa tattcgcaac gcgctgctgt tgcaagtccc attgacgggc
      721 tacattattg atagtaaggc gtttcttttc cttcaacagt gattgtttgt ggctttcgcg
      781 cttcttttcc tcctcttttt tagctaacac agacgccata cggcgctctt cctctttacg
      841 gtgctgcgtt gacggctgag gacagacgaa ctgaaacccc agcagtaata aaaaacacaa
      901 gacagacagg ctacccataa agtgcgacac catccgggaa gcggccgcac ttcccccgga
      961 gcctccgctt ccgcccttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
     1021 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
     1081 ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     1141 ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt
     1201 caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt
     1261 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     1321 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     1381 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     1441 gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag
     1501 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag
     1561 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     1621 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     1681 cttgctcacc atggtggcga ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat
     1741 tcggttcact aaacgagccc ggttcactaa acgagctctg cttatataga cctcccaccg
     1801 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     1861 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     1921 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     1981 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     2041 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     2101 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     2161 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     2221 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     2281 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     2341 agcacgcgtt cgaagcttga gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat
     2401 aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa aactattctt
     2461 aaacctacca agcctcctac tatcattatg aataatttta tataccacag ccaatttgtt
     2521 atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc ttgttcattc
     2581 ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta
     2641 atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag agatttatta
     2701 ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga gcaaccccaa
     2761 atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct tgcctggagc
     2821 tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat agccctcagc
     2881 aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac cgtcagcgtc
     2941 attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa caaagctcct
     3001 attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt gggtgctact
     3061 cctaatggtt caatttttac tactttatat ttatataatt cacttctcca attgtccctc
     3121 atatctcctc cttatggtgt tttactaaac ttttccatgt tctaatcctc atcctgtcta
     3181 cttgccacac aatcatcacc tgccatctgt tttccataat ccctaatgat ctttgctttt
     3241 cttcttggca ctacttttat gtcactatta tcttgtatta ctactgcccc ttcacctttc
     3301 cagaggagct ttgctggtcc tttccaaagt ggatttctgc tgtccctgta ataaacccga
     3361 aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt
     3421 atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg
     3481 tggatgaata ctgccatttg tactgctgtc ttaagatgtt cagcctgatc tcttacctgt
     3541 cctataattt tctttaattc tttattcata gattctacta ctccttgact ttggggattg
     3601 tagggaattt ccaggtctga agatcctgct gcttgctgtg cttttttctt acttttgttt
     3661 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     3721 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     3781 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     3841 gtattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     3901 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     3961 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     4021 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     4081 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     4141 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     4201 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     4261 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     4321 cagtacaggc aaaaagcagc tgcttatatg caggatctga gggctcgcca ctccccagtc
     4381 ccgcccaggc cacgcctccc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4441 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4501 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4561 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4621 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4681 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4741 taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg cacttttcgg
     4801 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4861 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4921 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4981 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5041 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5101 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5161 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5221 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5281 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5341 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5401 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5461 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5521 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5581 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5641 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5701 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5761 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5821 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5881 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6001 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6601 cctgcgttat cccctgcttc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6661 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6721 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag cggatccaga
     6781 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6841 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6901 acaagttagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6961 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     7021 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     7081 gactatggtt gctgactaat tgagatgcgg atccgctggc acgacaggtt tcccgactgg
     7141 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7201 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7261 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     7321 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     7381 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     7441 tat
//
