LOCUS       sequence-488149-        7380 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_126, derived from human fusion protein PAX3_FOXO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7380)
  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_126
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(482..501)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    547..602
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(601..620)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(963..979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(963..979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(976..998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    987..1003
                     /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)."
     promoter        complement(join(1011..1017,1018..1035,1036..1041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1056..1077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1136..1331
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1182..1317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1337..1471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1374..1393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1428..1447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1539..1556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1710..2298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1790..1809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2469..3260,3261..3329))
                     /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     3092..3111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3330..3434)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3465..3844
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3846..4103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4150..4275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4767..4884
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5244..5477
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5662..5706
                     /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"
     enhancer        5963..6266
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6267..6470
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6421..6441
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6546..6555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6552..6554,6555..6557,6558..7268)
                     /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(6597..6618)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6858..6877)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7205..7226
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaagc taagcacagt atcgacggta tcctgtcaga gcgtgcgtca gctccgcagt
       61 ccgacgaggg atcggatatt gacagcgaac ctgatctgcc gttaaagcgc aaacagcgcc
      121 gcagccgcac gacttttaca gctgaacagt tggaggaatt agagcgcgct tttgagcgca
      181 cccactaccc tgatatttac acgcgtgaag agctggcaca acgtgctaag tgaaccggtt
      241 ctagataaga ttacggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
      301 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag aagacaagat
      361 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
      421 ggctaactag ggaacccact gcttaagcct caataaagct tcagctgctc gagctagcag
      481 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
      541 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
      601 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
      661 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
      721 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
      781 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
      841 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
      901 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
      961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     1141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     1201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1321 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1441 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     1501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac
     3481 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     3541 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3601 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3661 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3721 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3781 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3841 catgggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     3901 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     3961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     4021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     4141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     4201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     4261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     4321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     4381 ctagaacgat tcgcagttaa tactggcctg ttagaaacat cagaaggctg tagacaaata
     4441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     4501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     4561 ttagacaaga tagaggaaga gcaaaacaaa agtaagaaaa aagcacagca agcagcagga
     4621 tcttcagacc tggaaattcc ctacaatccc caaagtcaag gagtagtaga atctatgaat
     4681 aaagaattaa agaaaattat aggacaggta agagatcagg ctgaacatct taagacagca
     4741 gtacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     4801 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     4861 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaaa tccactttgg
     4921 aaaggaccag caaagctcct ctggaaaggt gaaggggcag tagtaataca agataatagt
     4981 gacataaaag tagtgccaag aagaaaagca aagatcatta gggattatgg aaaacagatg
     5041 gcaggtgatg attgtgtggc aagtagacag gatgaggatt agaacatgga aaagtttagt
     5101 aaaacaccat aaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     5161 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     5221 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     5281 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     5341 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     5401 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     5461 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     5521 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     5581 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     5641 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     5701 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     5761 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     5821 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccg
     5881 agctcaagct tcgaacgcgt gctagttatt aatagtaatc aattacgggg tcattagttc
     5941 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6001 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6061 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6121 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6181 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6241 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     6301 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6361 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6421 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     6481 accgggctcg tttagtgaac cgaattctgc agtcgacggt accgcgggcc cgggatccac
     6541 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     6601 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     6661 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     6721 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     6781 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     6841 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     6901 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     6961 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     7021 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     7081 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     7141 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     7201 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     7261 tgtacaaggg cggaagcgga ggctccgggg gaagtgcggc cgcttcccgg ttgcgcagta
     7321 aattcggcaa aggggaggag gaagaggcgg atttagagcg taaggaggct gaagaatcag
//
