LOCUS       sequence-490661-        8679 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_017, derived from human fusion protein BCR_ABL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8679)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_017
FEATURES             Location/Qualifiers
     source          1..8679
                     /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(2212..2922,2923..2925,2926..2928))
                     /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(2254..2275)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2603..2622
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2862..2883
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2925..2934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3010..3213)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3039..3059)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3214..3517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3774..3818)
                     /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    4003..4236
                     /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    4596..4713
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5205..5330
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5377..5634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5636..6015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6046..6150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6151..6219,6220..7011)
                     /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(6369..6388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7182..7770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7671..7690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7924..7941
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    8009..8143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8033..8052)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8087..8106
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8149..8344)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8163..8298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8403..8424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8439..8444,8445..8462,8463..8469)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8477..8493
                     /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     8482..8504
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8501..8517
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8501..8517
                     /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 ggcgccgctg gcgatccgct ctggaggctg
      601 gcgggttttc cgaagagaca ctcgggttga tatgagaggg atgaaggcgg tggaagacgg
      661 ctggtcccct gccagggccg aggatgctga tggcaacgtg gagggaacgg gggctgggct
      721 gatgggggtc cccgacggct tggcggactg tggctttgga gtggccggga gcacgggctt
      781 tttgaggccc tctcccggct ggctgggctt ggcagcgtca ctgttcacag catcaaccag
      841 actcgtggct tttgtctttg cgcccaggac tgcctccccg gccgcctcct ggctcgggct
      901 ctgcgagggc tttcctccag ccttccctgc agaggctgct ggtgggggcg gcggggcagg
      961 tttgagcctg gacaatttcc ccttgtccct ccctggcgac tcagaggagt gcttgtgcct
     1021 cctcactctg gactcctcgg cggggccctt gctggtgccc cctggtgcac ctgagcctgc
     1081 tttgctggtg ggggtcactg gctcagctgc agcaggggtc cctaaggcac tgccctttcc
     1141 agcttcttcc ttgtggggga ggcccgaggc aggggccacg gtgacctgcc gccggagggg
     1201 ttttggagtc aggttgggcg ggctggagcc cgggctggac tccatgatgt ctttgaagac
     1261 ctcatcagca gcttcctcat tctttttcac cagcctgggg ggaggcgtta ctgtgcctcg
     1321 ggtcacctgg tcagacctgt tctcccctgc cctcttccga ggcagagccg gcttctcact
     1381 tttgtgccct ccaaatgtgg acgagtcaaa ctgtcttccc gtggactgca agtcccgagg
     1441 cagcgtgact gacctccact ccgtgtcctt ggccccatgg ggaacgcagg aggcggagca
     1501 agagcgcagg aagcgcttgc tggagctgcc accgccctcc tcctcgccgg tggctaggcg
     1561 gctgctggtc agcgtgctgg acttcttcca caggtgggga gaccggaagc ctgagccccc
     1621 ggactcccgg agggctccat tggggacccc agccccattg ctgggctttg gggacttggc
     1681 tgggtcagct gtgtccaagg gggtgaaagc cagtgccccg ttgctgatgt ctcggccctc
     1741 ttcctcgccg gcccctctgc gctccggctg gccgtccatc tcccggaagg agctgctgcg
     1801 tttgggaggg gttggggctg tcttcttctt cttcttgatc aaggcgctga acaagttggt
     1861 ctttttgtct ttggggagaa ggcgctcatc ttcattcagg ccgccctccg ggggacctcg
     1921 ctcttttcga gggagcaatg gagacacggc aggctcatgg tccagaggat cgctctctcc
     1981 ctggcccttg gagtgaggca tctcaggcac gtcagtggtg tctctgtgct ctgcagctct
     2041 cctggaggtc ctcgtcttgg tgggcagctc tggggcctgc agcaaggtac tcacagcccc
     2101 acggacgcct tgtttcccca gctccttttc cacttcgtct gagatactgg attcctggaa
     2161 cattgtttcc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc ccttgtacag
     2221 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     2281 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
     2341 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     2401 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     2461 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     2521 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     2581 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     2641 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     2701 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     2761 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     2821 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     2881 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     2941 tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac gagcccggtt
     3001 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     3061 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     3121 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     3181 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     3241 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3301 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     3361 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     3421 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     3481 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     3541 aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa gcttgagctc
     3601 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     3661 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     3721 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     3781 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     3841 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     3901 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     3961 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     4021 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     4081 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     4141 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     4201 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     4261 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     4321 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta tggtgtttta
     4381 ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc atcacctgcc
     4441 atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac ttttatgtca
     4501 ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc tggtcctttc
     4561 caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
     4621 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     4681 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtact
     4741 gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt taattcttta
     4801 ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag gtctgaagat
     4861 cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat cttgtctaaa
     4921 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     4981 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     5041 atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa tcgttctagc
     5101 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     5161 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     5221 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     5281 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     5341 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     5401 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     5461 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     5521 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     5581 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctcccatgg
     5641 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     5701 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     5761 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     5821 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     5881 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     5941 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     6001 tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6061 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6121 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6181 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6241 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6301 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6361 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6421 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6481 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6541 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6601 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6661 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6721 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6781 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6841 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6901 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6961 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7021 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7081 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7141 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7201 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7261 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7321 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7381 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7441 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7501 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7561 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7621 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7681 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7741 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7801 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgcttctgtg
     7861 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7921 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7981 gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca ttgatgagtt
     8041 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     8101 tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt tgcaaaagcc
     8161 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     8221 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     8281 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     8341 atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     8401 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     8461 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     8521 gattacgcca agcttgggct gcaggtcgag ggatctccat aagagaagag ggacagctat
     8581 gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt
     8641 tagggatgtt aaagaaaaaa ataacacaaa acaaaatat
//
