LOCUS       sequence-488212-        7713 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_193, derived from human fusion protein THBS1_FSIP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7713)
  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_193
FEATURES             Location/Qualifiers
     source          1..7713
                     /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(1246..1956,1957..1959,1960..1962))
                     /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(1288..1309)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1637..1656
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1896..1917
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1959..1968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2044..2247)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2073..2093)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2248..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2808..2852)
                     /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    3037..3270
                     /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    3630..3747
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4239..4364
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4411..4668
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4670..5049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5080..5184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5185..5253,5254..6045)
                     /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(5403..5422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6216..6804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6705..6724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6958..6975
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7043..7177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7067..7086)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7121..7140
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7183..7378)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7197..7332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7437..7458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7473..7478,7479..7496,7497..7503)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7511..7527
                     /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     7516..7538
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7535..7551
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7535..7551
                     /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 taactcttta ccctgatgac gaacaaccca
      601 gttaggatca ttctgggacg tgcctttcgg gtccagcgga atcatctgga agcgccgaaa
      661 atcggtttcg ctgatgtcca cgttctcagg gcagatgtcg tcgatgtccg gcaccgaatc
      721 gtgatcgaag tcatccttgc acgcatcccc acgaccgtcg ccgtcagagt ctttttgatc
      781 tgggttcggc accagacggc agttgtcctt atcgtccgga atgccatcgt tatcgtcatc
      841 gtggtcacac gcgtcgcctt taccatcctt gtcgtggtcg gcctggttcg cattcggaac
      901 gtacgggcag ttatccaagt tattctgatg gccatcttca tcgatatcct ggttgttgtc
      961 acaggtatcg ccgattcggt ctgagtccga atccagttgg tccgggttgt gctctaacgg
     1021 gcagttgtcg cattgatcac caacaccgtc catatcggtg tcacgctgat caacattata
     1081 gacgtactgg cagttgtcac gttcgttgag gatgccatca ccatcgatgt ccgctgcgca
     1141 cgcatcacct tcaccgttgt tgtcggtgtc tgcttggtcc ggattgtggt tgtacgggca
     1201 gttccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
     1261 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1321 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     1381 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1441 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1501 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1561 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1621 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1681 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1741 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1801 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1861 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1921 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     1981 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     2041 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     2101 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     2161 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     2221 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     2281 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     2341 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     2401 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2461 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2521 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2581 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2641 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2701 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2761 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2821 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     2881 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     2941 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     3001 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     3061 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     3121 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     3181 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     3241 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     3301 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     3361 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3421 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3481 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3541 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3601 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3661 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3721 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3781 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3841 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     3901 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     3961 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     4021 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     4081 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     4141 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     4201 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     4261 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     4321 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     4381 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4441 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4501 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4561 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4621 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4681 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4741 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4801 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4861 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4921 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4981 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     5041 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5401 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5701 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6841 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgcttc tgtggataac
     6901 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6961 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7021 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     7081 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     7141 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     7201 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     7261 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     7321 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     7381 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7441 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7501 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7561 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7621 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7681 tgttaaagaa aaaaataaca caaaacaaaa tat
//
