LOCUS       sequence-488174-        8520 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_153, derived from human fusion protein PAX7_FOXO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8520)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8520)
  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_153
FEATURES             Location/Qualifiers
     source          1..8520
                     /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(2053..2763,2764..2766,2767..2769))
                     /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(2095..2116)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2444..2463
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2703..2724
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2766..2775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2851..3054)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2880..2900)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3055..3358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3615..3659)
                     /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    3844..4077
                     /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    4437..4554
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5046..5171
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5218..5475
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5477..5856
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5887..5991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5992..6060,6061..6852)
                     /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(6210..6229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7023..7611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7512..7531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7765..7782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7850..7984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7874..7893)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7928..7947
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7990..8185)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8004..8139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8244..8265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8280..8285,8286..8303,8304..8310)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8318..8334
                     /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     8323..8345
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8342..8358
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8342..8358
                     /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 tcccagcgca ctcatctgca tagggtgggg
      601 taacggcacc tggactggag ttttgacttg ggtcaaacga ttcatccccg atgtgtgtgg
      661 catggtggat actgtatggg gtaacgggcg cccattcaca gctgaggtct gctgtgcgtg
      721 tccggggtgc gtgtgtgaac tcggattcat cattttattg tgggaagcct ggctaccata
      781 cgtcgacata acggagttgg gacccatcat aacattttga cccaatacgc gtgaattagg
      841 ttgggcgaca ccagggtcaa ccggcgtcat gatatcgttg tggggcgggg agtcagatgt
      901 aagcagttct ttcaacagcc caggcgcgca gttatattga ctcatgccac cgtaggaact
      961 cttattgtcc tgcagggttt ggattggcat ctgaggcaaa ggggacattg aggactgacc
     1021 ataagtgtac ttttgatagt tcggtgatgg agaattaagt gacgtatttg ggggggcgaa
     1081 actataacag ggggtttgct gcatcatcgt ccccggagag gattgcgttg acacggtcaa
     1141 ggaagtcgga gatgacagca ggtttaagtt gtctaacagg ttttccatat tttccggatt
     1201 agaaatctca cttaacgacg gaagagtaga cgccattttg gccgcggaag ggggatacac
     1261 catcgaatgc acgtcgcctt cgccaagatc atcctgctcc gtcatgatag gactcaagcg
     1321 accagaaata gtagaagcgt ttgatgaggt acgcggacgg aaggtagacc agttatcgaa
     1381 gtcgtcattg gaatgagaac ctggggaggc cggccactta ctaaattggc ttccagggct
     1441 atcgccagct ccttcttggc ccgattgaag tgatgccttc tttttggcgg cgcgcgaacg
     1501 ggatttggca aacttggaat tattatccat agatgctgca cgacgacgcg ggcttttccc
     1561 agatttgcca ccctcggggt tcaacatcca ccaagacgac ttcccagtcc cctcattttg
     1621 aacacggatg aatttggaat gaaggcttaa attatggcgg atggaatttt gaggggataa
     1681 tccattactg acggggttca tgtgattact aggtgccgca gggttcatga agctgtcact
     1741 ataagatgag aaagagtgac gagccccata cgcgcttgat gtatctgcgg cggcagcagc
     1801 ggcggccaat ccaccctggt gcatagtaga gggcggtaaa ggctgtggac gatgcactgt
     1861 ggagccacca tcctgagaga ttgtcgttgt agggtaggtg gagtccggta actgataggg
     1921 aggcaatgtc ggcatacctg taggtggaaa tcccccaggc agaagatgat tgaatgctgc
     1981 cagctgattg gccccggctt gtttacgcca ccgggaagcg gccgcacttc ccccggagcc
     2041 tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2101 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt
     2161 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2221 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     2281 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     2341 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2401 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2461 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2521 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     2581 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2641 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2701 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2761 gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact gcagaattcg
     2821 gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct cccaccgtac
     2881 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     2941 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     3001 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     3061 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     3121 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     3181 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     3241 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     3301 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     3361 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagc
     3421 acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg aaacgataat
     3481 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     3541 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     3601 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     3661 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     3721 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     3781 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     3841 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     3901 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     3961 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     4021 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     4081 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     4141 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     4201 tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc ctgtctactt
     4261 gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt tgcttttctt
     4321 cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc acctttccag
     4381 aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata aacccgaaaa
     4441 ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt tgctattatg
     4501 tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt aaaattgtgg
     4561 atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct tacctgtcct
     4621 ataattttct ttaattcttt attcatagat tctactactc cttgactttg gggattgtag
     4681 ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact tttgttttgc
     4741 tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca
     4801 caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg
     4861 gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagta
     4921 ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt
     4981 ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta
     5041 atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc
     5101 gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg
     5161 cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc
     5221 tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca
     5281 acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg
     5341 ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag
     5401 tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc cccagtcccg
     5461 cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     5521 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     5581 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     5641 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     5701 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     5761 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     5821 ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7681 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7741 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7801 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcgg atccagacat
     7861 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     7921 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     7981 agttagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     8041 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     8101 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     8161 tatggttgct gactaattga gatgcggatc cgctggcacg acaggtttcc cgactggaaa
     8221 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8281 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8341 acaggaaaca gctatgacca tgattacgcc aagcttgggc tgcaggtcga gggatctcca
     8401 taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag
     8461 taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat
//
