LOCUS       sequence-487934-        8328 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_019, derived from human fusion protein BCR_ABL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8328)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8328)
  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_019
FEATURES             Location/Qualifiers
     source          1..8328
                     /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(1861..2571,2572..2574,2575..2577))
                     /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(1903..1924)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2252..2271
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2511..2532
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2574..2583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2659..2862)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2688..2708)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2863..3166
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3423..3467)
                     /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    3652..3885
                     /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    4245..4362
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4854..4979
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5026..5283
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5285..5664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5695..5799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5800..5868,5869..6660)
                     /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(6018..6037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6831..7419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7320..7339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7573..7590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7658..7792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7682..7701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7736..7755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7798..7993)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7812..7947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8052..8073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8088..8093,8094..8111,8112..8118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8126..8142
                     /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     8131..8153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8150..8166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8150..8166
                     /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 ggactccagg gctcccgaga ccagcgcatc
      601 ccggctgccc ctgcccttgc tgctgaggtg gggcgatgag gagggcgagt catcaatgta
      661 gggcagccca tcttggtgcc ggcgctgctc ctctgcccgg tctgcagcgc agccgtatcc
      721 aggtggtgtt ccgaacgagc catctgcgtc tccatggaag gcgccctcgc catcgttggg
      781 ccagatctgc ccggtcttgc ggacgcccac gatggtggcc tcggacacga caaccgggca
      841 gtgccggtgc cgcttatggc actgcggcgt ggggggactg ctgctgtcga aggactgttg
      901 cgagttctgc gagggagagc ggcttttgtc ccggaacatg cggtaggtgg tggggcttgg
      961 ggacacgcgg ctggactggc cagaggagaa gtcctcctcg ctggaggtga ggttctcatt
     1021 ggagctgcag tccggggtat agccgcctcc gcaatcctca aaactccggg gggagtagga
     1081 cctgcggggc caggtaaggc gcttctcctg ctcagaggtg ctctggctgc gcaggagcgg
     1141 gcccttgccc tccccttcca tcatgccccc gacgtagatg ctctggtagg gctggtactc
     1201 caggggcggc caagggggcc tgctaccgcc attggcgtcg atcaggttgt ccttcaggaa
     1261 gcgggggttc aactcggcgt cctcgtagtc gccgtcgacg ccgcagctgc tctccgagga
     1321 gcgtccccgg taagggggcc tggatgcatc ccccacgctc gagcccgcgc cgtgctggga
     1381 ctttttgcgc tccatctgca tggcctggct gcccagggag ctgatgcggt ccgacacctc
     1441 tttgtcgttg accttcacca ggccgcgctc gtggtgaaac tcgacgttca cgtagaaggg
     1501 cttctcggcg tccgccccgg gctggccatg gcccttgcgg atccgctcga agttggacct
     1561 gagcgccgcc acgctggcgg ggggtccccg gtcgtcccgt tcccccgacg cggctgcccc
     1621 gggcctgcgg gcggtcccgg gcctggcctt acccggagaa ccctcgccgt cgggccgggc
     1681 ctcgggctcc tcggcgggcg gcgggtcggc tccgtcggcg ggcgctggct gcgggcgcga
     1741 cgcggacgct cggggctcgg aggcgccgtc gggggcctgc gccgcgcgcc ggaagcccca
     1801 tcgctgccgg tcatagctcc gggaagcggc cgcacttccc ccggagcctc cgcttccgcc
     1861 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     1921 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc tcaggtagtg
     1981 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     2041 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     2101 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     2161 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     2221 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     2281 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     2341 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     2401 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     2461 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     2521 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     2581 ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt tcactaaacg
     2641 agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     2701 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     2761 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     2821 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     2881 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     2941 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     3001 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     3061 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     3121 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     3181 tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac gcgttcgaag
     3241 cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc
     3301 tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct
     3361 cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc
     3421 cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt
     3481 tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac
     3541 tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc
     3601 caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt
     3661 tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag
     3721 actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc
     3781 tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc
     3841 atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt
     3901 ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt
     3961 tttactactt tatatttata taattcactt ctccaattgt ccctcatatc tcctccttat
     4021 ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc cacacaatca
     4081 tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct tggcactact
     4141 tttatgtcac tattatcttg tattactact gccccttcac ctttccagag gagctttgct
     4201 ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt
     4261 gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt
     4321 tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc
     4381 atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat aattttcttt
     4441 aattctttat tcatagattc tactactcct tgactttggg gattgtaggg aatttccagg
     4501 tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc ttcctctatc
     4561 ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca atagagggtt
     4621 gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga tggttgtagc
     4681 tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt aactgcgaat
     4741 cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt ctttccccct
     4801 ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat actgacgctc
     4861 tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt actcaccagt
     4921 cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc
     4981 tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta gagattttcc
     5041 acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac
     5101 acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt
     5161 agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta caggcaaaaa
     5221 gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc caggccacgc
     5281 ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     5341 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     5401 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     5461 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     5521 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     5581 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     5641 aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga
     5701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6001 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6841 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6901 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6961 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7021 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7081 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7141 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7201 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7261 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7321 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7381 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7441 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7501 gcttctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7561 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7621 cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga taagatacat
     7681 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     7741 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttagcttttt
     7801 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     7861 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     7921 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     7981 ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8041 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8101 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8161 tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata agagaagagg
     8221 gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta aaacatgtaa
     8281 ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatat
//
