LOCUS       sequence-487974-        7671 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_064, derived from human fusion protein ETV6_NTRK3; ETV6_RUNX1;
            PAX5_ETV6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7671)
  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_064
FEATURES             Location/Qualifiers
     source          1..7671
                     /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(1204..1914,1915..1917,1918..1920))
                     /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(1246..1267)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1595..1614
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1854..1875
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1917..1926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2002..2205)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2031..2051)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2206..2509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2766..2810)
                     /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    2995..3228
                     /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    3588..3705
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4197..4322
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4369..4626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4628..5007
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5038..5142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5143..5211,5212..6003)
                     /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(5361..5380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6174..6762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6663..6682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6916..6933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7001..7135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7025..7044)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7079..7098
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7141..7336)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7155..7290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7395..7416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7431..7436,7437..7454,7455..7461)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7469..7485
                     /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     7474..7496
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7493..7509
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7493..7509
                     /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 gcgaccgatt ggcatggcat gttcctcggg
      601 ggggcttacc gaaaccatga tatggttcat ataagcaagg tcctcacgat gtgacaagtt
      661 aattggcttt ccttcacggt gaagtccatc ctctgacaag cgactttgct tgaagtctac
      721 ggaatggcgc gggttaagga tcaatgggtg cataatagga gacggcataa gttggattac
      781 acgcgtactt tcttggcgtg gactgcttgg cttggggtgg ctctctgacg aggcagggca
      841 atggttgttt tccatgggag aaacgctcaa cggataactt tcttggtgat tgttttcttg
      901 atgcggacgg gggccttgcg cgcgttcggc cgggctcaga cgacggatca tgttgtctaa
      961 gggactacgc agtggacgtt gctccgggtc gggtgaagga cggtggtttg tcgtgattgg
     1021 gctacggcta cggtgaagca gttcgatggt aggtggattg tgatgtacgt tgtcaacgga
     1081 agggcgtggg gtgcgttgta cgcaattgtc ctcttcgtga ttctggtgta aaattacctc
     1141 tggttgtgtg tggatggagt tccgggaagc ggccgcactt cccccggagc ctccgcttcc
     1201 gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     1261 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg tgctcaggta
     1321 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     1381 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
     1441 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
     1501 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1561 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1621 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1681 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1741 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1801 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1861 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1921 ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc ggttcactaa
     1981 acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc
     2041 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     2101 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     2161 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     2221 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     2281 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     2341 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     2401 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     2461 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     2521 atatgggcta tgaactaatg accccgtaat tgattactat taataactag cacgcgttcg
     2581 aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa tggtgaatat
     2641 ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa acctaccaag
     2701 cctcctacta tcattatgaa taattttata taccacagcc aatttgttat gttaaaccaa
     2761 ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt ttcttgctgg
     2821 ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc
     2881 cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact ccaactagca
     2941 ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat ccccaggagc
     3001 tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg cttgatgccc
     3061 cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa attgttctgc
     3121 tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg
     3181 cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat tcccactgct
     3241 cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc taatggttca
     3301 atttttacta ctttatattt atataattca cttctccaat tgtccctcat atctcctcct
     3361 tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact tgccacacaa
     3421 tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct tcttggcact
     3481 acttttatgt cactattatc ttgtattact actgcccctt cacctttcca gaggagcttt
     3541 gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa attttgaatt
     3601 tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt
     3661 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact
     3721 gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc tataattttc
     3781 tttaattctt tattcataga ttctactact ccttgacttt ggggattgta gggaatttcc
     3841 aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg ctcttcctct
     3901 atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg
     3961 gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt
     4021 agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt attaactgcg
     4081 aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt tttctttccc
     4141 cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg
     4201 ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc
     4261 agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag
     4321 agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt
     4381 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     4441 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     4501 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa
     4561 aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca
     4621 cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     4681 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     4741 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     4801 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     4861 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     4921 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4981 attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg aaatgtgcgc
     5041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5161 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5281 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5341 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5521 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5701 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5761 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5821 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6001 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6841 cctgcttctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6901 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6961 ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg gatccagaca tgataagata
     7021 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     7081 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttagctt
     7141 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     7201 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     7261 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     7321 tgactaattg agatgcggat ccgctggcac gacaggtttc ccgactggaa agcgggcagt
     7381 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7441 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7501 agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc ataagagaag
     7561 agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta gtaaaacatg
     7621 taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata t
//
