LOCUS       sequence-487973-        7599 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_063, 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 7599)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7599)
  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_063
FEATURES             Location/Qualifiers
     source          1..7599
                     /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(1132..1842,1843..1845,1846..1848))
                     /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(1174..1195)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1523..1542
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1782..1803
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1845..1854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1930..2133)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1959..1979)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2134..2437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2694..2738)
                     /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    2923..3156
                     /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    3516..3633
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4125..4250
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4297..4554
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4556..4935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4966..5070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5071..5139,5140..5931)
                     /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(5289..5308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6102..6690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6591..6610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6844..6861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6929..7063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6953..6972)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7007..7026
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7069..7264)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7083..7218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7323..7344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7359..7364,7365..7382,7383..7389)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7397..7413
                     /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     7402..7424
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7421..7437
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7421..7437
                     /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 gtgagacagg ttaattggtt ttccctcacg
      601 gtgaaggcca tcctcactca ggcgggactg tttaaagtct actgagtgac gagggttaag
      661 gataagaggg tgcatgattg gactaggcat taactggatc acgcgcgtcg attcttggcg
      721 aggagacgaa ggtttagggt gagactcaga actggctggg caatggttgt tttccatagg
      781 ggagacagac agtgggtagg attcctggtg attattctcc tgatgggggc gtgggccttg
      841 ggcgcgttcc gccggagaca aacgacggat catgttgtcc aatggagagc gcaaggggcg
      901 ctgttccggg tcgggagacg ggcgatgatt tgttgtaatg gggctgcgtg agcggtgcaa
      961 aagttcgata gtgggtgggt tgtgatgaac gttatcgaca ctgggacggg gagtgcgttg
     1021 aacacagtta tcttcttcgt gattttgatg caggatgacc tcgggctgag tatggatcga
     1081 atttccgggc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc ccttgtacag
     1141 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     1201 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
     1261 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     1321 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     1381 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     1441 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     1501 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     1561 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     1621 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     1681 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     1741 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     1801 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     1861 tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac gagcccggtt
     1921 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     1981 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2041 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2101 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2161 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2221 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2281 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2341 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2401 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2461 aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa gcttgagctc
     2521 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     2581 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     2641 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     2701 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     2761 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     2821 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     2881 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     2941 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     3001 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     3061 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     3121 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     3181 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     3241 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta tggtgtttta
     3301 ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc atcacctgcc
     3361 atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac ttttatgtca
     3421 ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc tggtcctttc
     3481 caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
     3541 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     3601 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtact
     3661 gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt taattcttta
     3721 ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag gtctgaagat
     3781 cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat cttgtctaaa
     3841 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     3901 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     3961 atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa tcgttctagc
     4021 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     4081 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     4141 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     4201 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     4261 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     4321 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     4381 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     4441 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     4501 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctcccatgg
     4561 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4621 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4681 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4741 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4801 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4861 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4921 tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4981 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5041 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5101 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5161 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5221 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5281 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5341 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5401 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5461 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5521 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5581 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5641 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5701 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5761 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5821 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5881 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5941 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6001 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6061 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6241 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6721 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgcttctgtg
     6781 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6841 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6901 gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca ttgatgagtt
     6961 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     7021 tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt tgcaaaagcc
     7081 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     7141 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     7201 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     7261 atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7321 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7381 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7441 gattacgcca agcttgggct gcaggtcgag ggatctccat aagagaagag ggacagctat
     7501 gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt
     7561 tagggatgtt aaagaaaaaa ataacacaaa acaaaatat
//
