LOCUS       sequence-488127-        8250 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_102, derived from human fusion protein MED13L_COL23A1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8250)
  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_102
FEATURES             Location/Qualifiers
     source          1..8250
                     /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(1783..2493,2494..2496,2497..2499))
                     /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(1825..1846)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2174..2193
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2433..2454
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2496..2505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2581..2784)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2610..2630)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2785..3088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3345..3389)
                     /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    3574..3807
                     /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    4167..4284
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4776..4901
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4948..5205
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5207..5586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5617..5721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5722..5790,5791..6582)
                     /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(5940..5959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6753..7341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7242..7261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7495..7512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7580..7714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7604..7623)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7658..7677
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7720..7915)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7734..7869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7974..7995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8010..8015,8016..8033,8034..8040)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8048..8064
                     /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     8053..8075
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8072..8088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8072..8088
                     /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 tttatgccag caccccggta ccggcaagcc
      601 atccggaccc accgggcacg gttggtccaa gcccggaggc cccggttcgc ctttttgacc
      661 tttgacacct ttgcggcccg gcacaccgcg ctcgcccttt tcgccacgct cgctacgatc
      721 gcctttttcg ccccgcgggc ccggaaaacc gtccaggcct ggctcgcccg gacggccctt
      781 ctcgcctttg gtccccggca agccaatcaa gccaggcgga cccaccggtc ccggcaaacc
      841 ggacggacca cgttcaccgg acgcgccctt ttctcctttg gcaccgtcta ggcctttcgg
      901 tccctggata ccctgcaaac ccatgggacc cggcggcccc ggtgggcccg ggggtcccgg
      961 ttcaacaatc agctgtgcca gagattcttg caggctatca cttgcgcttt cccccttctc
     1021 acccttcagc ccatccgcac ccggcagacc agagagaccc atctcacccg cttccccctt
     1081 aagaccggca ggacccggtt cacccggatc acctttttgg cccttcggac ccttttcacc
     1141 atcgatacca ggcgcacccg gcagacccag ctcacctttc gcgcccggga tgcccggtgg
     1201 ccccggcgga ccctgcgggc ccggcggccc tttcagtgca tccagaatac ggccgtcgta
     1261 gtcgataaca accgtgtcac cctgctcgcc ctttaagccc ggagcaccgc gaggacccgc
     1321 cgccccgtcg gtaccacggt ggcccggctc accctttggg cctggcgcgc catcaacacc
     1381 gttctcacct ctcggaccca tgctgcccgg ctcacctttc ggacccggcg ggcccggctg
     1441 gctcggagtg ccatcgtcgc cttttttccc cggcacgccc ggttcgccct taggaccagg
     1501 cgggcccttc gggcccatct caccgtcttg gcccggctca cccttcgggc ctgctgggcc
     1561 ctgcgcaccg cgcggtccat ccttaccggt gtcgccgggc ggaccacgcg cccccggtgg
     1621 acccggcggg cccggcggac cggcagcgcc atcttgaccc tggtcaccca ggctgaacag
     1681 attgctgtga caatcttcca gcgaagcgcc atttgctacc cagttcgcag cagctgtcat
     1741 ccgggaagcg gccgcacttc ccccggagcc tccgcttccg cccttgtaca gctcgtccat
     1801 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     1861 gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac
     1921 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     1981 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     2041 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     2101 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2161 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     2221 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2281 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     2341 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     2401 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2461 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
     2521 ggcccgcggt accgtcgact gcagaattcg gttcactaaa cgagcccggt tcactaaacg
     2581 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     2641 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     2701 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     2761 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     2821 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     2881 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     2941 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     3001 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     3061 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     3121 ccccgtaatt gattactatt aataactagc acgcgttcga agcttgagct cggtcccctc
     3181 ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac tctattcact
     3241 atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat cattatgaat
     3301 aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact tgcccattta
     3361 tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc ttcaattaag
     3421 gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca ggtcgtgtga
     3481 ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac agcagtggtg
     3541 caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt taggtatctt
     3601 tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag ttgcaacaga
     3661 tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat accagacaat
     3721 aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct
     3781 cccaagaacc caaggaacaa agctcctatt cccactgctc ttttttctct ctgcaccact
     3841 cttctctttg ccttggtggg tgctactcct aatggttcaa tttttactac tttatattta
     3901 tataattcac ttctccaatt gtccctcata tctcctcctt atggtgtttt actaaacttt
     3961 tccatgttct aatcctcatc ctgtctactt gccacacaat catcacctgc catctgtttt
     4021 ccataatccc taatgatctt tgcttttctt cttggcacta cttttatgtc actattatct
     4081 tgtattacta ctgccccttc acctttccag aggagctttg ctggtccttt ccaaagtgga
     4141 tttctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     4201 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     4261 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtac tgctgtctta
     4321 agatgttcag cctgatctct tacctgtcct ataattttct ttaattcttt attcatagat
     4381 tctactactc cttgactttg gggattgtag ggaatttcca ggtctgaaga tcctgctgct
     4441 tgctgtgctt ttttcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     4501 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     4561 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     4621 cagccttctg atgtttctaa caggccagta ttaactgcga atcgttctag ctccctgctt
     4681 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     4741 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     4801 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     4861 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     4921 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     4981 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     5041 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     5101 ctcagatctg gtctaaccag agagacccag tacaggcaaa aagcagctgc ttatatgcag
     5161 gatctgaggg ctcgccactc cccagtcccg cccaggccac gcctcccatg gtaatagcga
     5221 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     5281 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     5341 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     5401 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     5461 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     5521 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc
     5581 aatgtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5641 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5701 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5761 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5821 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5881 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5941 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6001 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6061 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6121 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6181 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6241 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6301 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6361 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6421 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6481 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6541 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6601 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6661 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6721 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6781 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6841 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6901 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6961 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7021 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7081 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7141 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7201 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7261 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7321 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7381 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgcttctgt ggataaccgt
     7441 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7501 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7561 ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt ttggacaaac
     7621 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     7681 atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc ctaggcctcc
     7741 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     7801 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     7861 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcggatc
     7921 cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7981 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     8041 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     8101 aagcttgggc tgcaggtcga gggatctcca taagagaaga gggacagcta tgactgggag
     8161 tagtcaggag aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt
     8221 taaagaaaaa aataacacaa aacaaaatat
//
