LOCUS       sequence-488150-        8517 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_127, derived from human fusion protein PAX3_FOXO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8517)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8517)
  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_127
FEATURES             Location/Qualifiers
     source          1..8517
                     /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(2050..2760,2761..2763,2764..2766))
                     /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(2092..2113)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2441..2460
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2700..2721
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2763..2772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2848..3051)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2877..2897)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3052..3355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3612..3656)
                     /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    3841..4074
                     /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    4434..4551
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5043..5168
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5215..5472
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5474..5853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5884..5988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5989..6057,6058..6849)
                     /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(6207..6226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7020..7608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7509..7528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7762..7779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7847..7981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7871..7890)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7925..7944
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7987..8182)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8001..8136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8241..8262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8277..8282,8283..8300,8301..8307)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8315..8331
                     /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     8320..8342
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8339..8355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8339..8355
                     /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 tggggtgtgg
      601 cagcggtact tggaccgggg tctttacctg ggtcaagcga ttcattccag aagtatgggg
      661 cattgttgag acggtatggg gtaacgggcg accattgaca gcggatgttt gttgcgcatg
      721 acccggatga gtgtgagaac tcggattcat cattttattg tgtgaagcct gactaccgta
      781 agttgacatc acagaattag gccccatcat gacgttttgc cctaaaacac gactgtttgg
      841 ctgtgcaact ccaggatcaa cgggagtcat aatatcattg tgagggggtg aatccgaagt
      901 caacaactct ttcaaaaggc ccggggcgca attatattgt gacatcccgc cgtatgagga
      961 tttattgtct tgtaaggtct ggatgggcat ctgtggcaac ggcgacattg aactttgccc
     1021 gtatgtgtat ttctgatagt tcggactcgg actattaaga gaagtattag gcggagcaaa
     1081 agaataacag ggcgtctgct gcatcatggt gcctggagac gattgggttg atactgttaa
     1141 cgatgtgggg gaagacaaca aattcagatt gtccaataag ttctccatat tttccgggtt
     1201 actgatctct gacagtgaag gcaaagtaga ggccatcttg gctgcagagg ggggataaac
     1261 catagagtga acgtcgcctt cgcccaaatc gtcttgttca gtcatgattg gagacaaacg
     1321 gccgctgatt gtgctggcat ttgaactagt gcgggggcga aatgtgctcc agttatcgaa
     1381 atcgtcattc gagtgacttc ctggactcgc aggccacttg ctgaactggg acccgggaga
     1441 gtcgcctgca ccctcctgac cagactgcag gctagccttt ttcttagcgg cacgagaacg
     1501 gctctttgca aatttagagt tgttatccat tgaggcggca cgacggcgag gggacttacc
     1561 actcttgcca ccttcaggat taagcatcca ccacgatgat tttcctgtac cttcgttctg
     1621 tacacgaata aatttactat gtaagctcaa attatgacga atgctattct ggggtgacag
     1681 gccgttacca atcgtgggat tcatagggtt tgagggacca gaaggtggaa cgaagctgtc
     1741 agtgtagctg gaaaaaccat ggcgtgtgct gggaagacag tacgccgaac tagagtctgg
     1801 gttgctagga atcgtggatt gatgaactgt agacggtggt aagggctgag gacgatgcac
     1861 agtcgaactg ggatcagata ccgcttgggg gatgcttgtg ggctgatatg acgtctcact
     1921 caattggtac gtgggcagag tgggcatcgc ggtcgggggg aaaccaccag ggatcaaatg
     1981 gttaaaagcc attaattggt tcgcgccccg ggaagcggcc gcacttcccc cggagcctcc
     2041 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     2101 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     2161 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     2221 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     2281 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     2341 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     2401 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     2461 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     2521 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     2581 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     2641 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     2701 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2761 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     2821 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2881 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2941 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     3001 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     3061 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     3121 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     3181 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     3241 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     3301 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3361 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     3421 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     3481 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     3541 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     3601 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     3661 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     3721 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     3781 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     3841 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     3901 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     3961 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     4021 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     4081 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     4141 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     4201 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     4261 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     4321 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     4381 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     4441 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     4501 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     4561 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     4621 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     4681 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     4741 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     4801 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     4861 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     4921 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     4981 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     5041 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     5101 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     5161 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     5221 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     5281 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     5341 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     5401 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     5461 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     5521 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     5581 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     5641 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     5701 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     5761 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5821 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     5881 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5941 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6001 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6061 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6121 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6181 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6241 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6301 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6361 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6421 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6481 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6541 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6601 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6661 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6721 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6781 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6841 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6901 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7081 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7141 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7321 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7381 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7441 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7501 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7561 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7621 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7681 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7741 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7801 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     7861 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     7921 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     7981 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     8041 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     8101 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     8161 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     8221 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8281 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8341 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     8401 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     8461 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
