LOCUS       sequence-490662-        8058 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_018, derived from human fusion protein BCR_ABL1; BCR_JAK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8058)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8058)
  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_018
FEATURES             Location/Qualifiers
     source          1..8058
                     /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             951..965
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     CDS             complement(join(1591..2301,2302..2304,2305..2307))
                     /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(1633..1654)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1982..2001
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2241..2262
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2304..2313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2389..2592)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2418..2438)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2593..2896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3153..3197)
                     /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    3382..3615
                     /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    3975..4092
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4584..4709
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4756..5013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5015..5394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5425..5529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5530..5598,5599..6390)
                     /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(5748..5767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6561..7149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7050..7069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7303..7320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7388..7522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7412..7431)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7466..7485
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7528..7723)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7542..7677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7782..7803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7818..7823,7824..7841,7842..7848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7856..7872
                     /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     7861..7883
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7880..7896
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7880..7896
                     /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 atgacggtga cgtttgtggc attgaggtgt
      601 tggtggacta gatgagtcga aactttgctg gctattttga gacggggaac ggcttttatc
      661 gcggaacatg cgatacgtcg taggagatgg ggaaacacgg ctagactgtc cactagaaaa
      721 atcctcttca ctagacgtaa ggttttcgtt tgagctacaa tcgggcgtat agccaccgcc
      781 gcagtcttcg aagctacgag gggagtatga gcggcgcggc cacgtcaggc gcttttcttg
      841 ctcagaggtg ctctgggagc gcagaagtgg ccccttcccc tcaccttcca tcataccgcc
      901 cacgtagata ctctggtacg gttgatactc cagcggtggc cagggtgggc gggagccacc
      961 attggcgtcg atcaggttat ctttcaggaa gcgcggatta agctctgcat cttcgtagtc
     1021 accatcgaca ccgcagctag actcggatga gcgcccgcga tagggggggc ggctcgcatc
     1081 accaacggag ctaccggcgc catgctggga cttcttacgt tccatttgca ttgcttggct
     1141 acccaaactg ctgatacgat cggaaacctc tttatcattg accttaacaa gcccgcgttc
     1201 atgatgaaat tcaacgttaa cgtaaaacgg cttttccgcg tcagcaccgg gttgcccatg
     1261 tcctttacgg atgcgttcga aattgctgcg cagagcagct acgctcgcgg gaggcccacg
     1321 gtcatcgcgc tcaccagagg ctgctgcgcc agggcgacgc gcggtgccag ggcgagcttt
     1381 gccaggagaa ccttccccat ctggacgggc ttccggctcc tctgctggag ggggatcggc
     1441 tccatcggcg ggggccggtt gcggacgact tgctgaggca cgtggctcac tggctccatc
     1501 gggcgcttga gccgcacggc gaaagcccca acgttgacga tcataagacc gggaagcggc
     1561 cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat
     1621 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     1681 gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1741 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
     1801 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
     1861 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1921 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     1981 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     2041 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     2101 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     2161 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     2221 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     2281 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg cccgcggtac
     2341 cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag ctctgcttat
     2401 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     2461 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     2521 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     2581 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     2641 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     2701 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2761 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2821 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2881 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2941 ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg
     3001 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     3061 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     3121 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     3181 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     3241 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     3301 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     3361 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     3421 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     3481 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     3541 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     3601 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     3661 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     3721 ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc catgttctaa
     3781 tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc ataatcccta
     3841 atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg tattactact
     3901 gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt tctgctgtcc
     3961 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     4021 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     4081 tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag atgttcagcc
     4141 tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc tactactcct
     4201 tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg ctgtgctttt
     4261 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     4321 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     4381 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     4441 gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     4501 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     4561 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     4621 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     4681 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     4741 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     4801 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     4861 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     4921 ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct
     4981 cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg actaatacgt
     5041 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     5101 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     5161 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     5221 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     5281 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     5341 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgaccca
     5401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5521 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5581 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5641 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5701 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5761 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5821 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5881 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5941 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6001 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6061 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6121 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6181 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6241 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6301 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6361 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6421 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6481 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7201 gctcacatgt tctttcctgc gttatcccct gcttctgtgg ataaccgtat taccgccttt
     7261 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7321 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7381 tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     7441 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7501 tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     7561 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     7621 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     7681 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac
     7741 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7801 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7861 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg
     7921 caggtcgagg gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     7981 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     8041 taacacaaaa caaaatat
//
