LOCUS       sequence-488175-        7353 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_155, derived from human fusion protein PIEZO1_CBFA2T3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7353)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7353)
  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_155
FEATURES             Location/Qualifiers
     source          1..7353
                     /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(886..1596,1597..1599,1600..1602))
                     /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(928..949)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1277..1296
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1536..1557
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1599..1608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1684..1887)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1713..1733)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1888..2191
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2448..2492)
                     /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    2677..2910
                     /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    3270..3387
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3879..4004
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4051..4308
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4310..4689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4720..4824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4825..4893,4894..5685)
                     /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(5043..5062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5856..6444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6345..6364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6598..6615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6683..6817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6707..6726)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6761..6780
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6823..7018)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6837..6972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7077..7098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7113..7118,7119..7136,7137..7143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7151..7167
                     /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     7156..7178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7175..7191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7175..7191
                     /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 acacgcaggg ggtaaatgtt gggtagacag
      601 ggaagcggta ctcgaggtcg ccggcccgtt cgaaaagccg ttgggtgtac agggcgcgcc
      661 attgatagcc gtcggactgt gcgaagatcc gttcataagg gtgtgcggtg taaatgaggg
      721 agggcgagta gctccttgag aggccgccgg gggagggggt ggcatactgg gcggggtact
      781 gcgaggctga gtttttactt cagctgggct gtctggcata gcactcgctt tagctttgcg
      841 atcccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
      901 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
      961 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     1021 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1081 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1141 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1201 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1261 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1321 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1381 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1441 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1501 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     1561 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     1621 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     1681 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     1741 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     1801 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     1861 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     1921 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     1981 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     2041 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2101 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2161 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2221 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2281 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2341 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2401 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2461 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     2521 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     2581 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     2641 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     2701 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     2761 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     2821 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     2881 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     2941 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     3001 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3061 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3121 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3181 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3241 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3301 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3361 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3421 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3481 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     3541 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     3601 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     3661 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     3721 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     3781 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     3841 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     3901 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     3961 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     4021 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4081 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4141 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4201 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4261 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4321 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4381 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4441 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4501 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     4561 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     4621 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     4681 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4741 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4801 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4861 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4921 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4981 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5041 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5101 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5161 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5221 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5281 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5341 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5401 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5461 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5521 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5581 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5641 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5701 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5761 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5821 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5881 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5941 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6001 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6061 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6121 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6181 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6241 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6301 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6361 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6421 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6481 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6541 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6601 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6661 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     6721 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6781 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     6841 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     6901 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     6961 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     7021 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7081 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7141 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7201 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7261 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7321 tgttaaagaa aaaaataaca caaaacaaaa tat
//
