LOCUS       sequence-267782-        6363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A reporter construct containing the Exon 5 of PTPRC (CD45), which 
            contains a mutant CTCF binding site, making the exon to be always 
            excluded..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6363)
  AUTHORS   Wang AJ, Han Y, Jia N, Chen P, Minden MD
  TITLE     NPM1c impedes CTCF functions through cytoplasmic mislocalization in 
            acute myeloid leukemia.
  JOURNAL   Leukemia. 2020 May;34(5):1278-1290. doi: 10.1038/s41375-019-0681-8. 
            Epub 2019 Dec 12.
  PUBMED    31831844
REFERENCE   2  (bases 1 to 6363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41375-019-0681-8"; journalName: "Leukemia"; date: 
            "2020-05"; volume: "34"; issue: "5"; pages: "1278-1290"
COMMENT     Sequence Label: pFlare9A-CD45i4e5i5mutCBS
FEATURES             Location/Qualifiers
     source          1..6363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1445..1454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1590..1592,1593..2306)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHLKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGTDELYK"
     primer_bind     complement(1632..1653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1893..1912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2313..2322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2319..3029
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2464..2482)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2732..2751
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3151..3272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3188..3207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3242..3261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3279..3734)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3416..3437)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3628..3647
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3761..3865
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3867..4224
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3867..4058
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3890..3910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4075..4210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4259..5053
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4259..5053
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4313..4332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4923..4942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5241..5260)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5285..5332
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5661..6249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6150..6169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctaccatt ggtgcacctg actcctgagg agaagtctgc cgttactgcc
      661 ctgtggggca aggtgaacgt ggaagagttg gtggtgaggc cctgggccac cagtaagtat
      721 caaggttaca agacaggttt aaggagacca atagaaactg ggcatgtgga gacagagaag
      781 actcacgcgt ttctgaattc tctatggctg catccagctc aggaaacctc ccaaggctat
      841 gtaaatagag gactcaggtg cctgtcactt tccagccaag caaatttaaa tactgacaga
      901 cacattttaa cagattcagt tacttcacag tttggtaaac tataatcgca atttgctcct
      961 tataacctct tagtaaatta ttcatctcca gtgggggaag actgatgtaa tgatctcact
     1021 ttcctacctt aggtgtttca tcagtacaga cgcctcacct tcccacgcac gcagactcgc
     1081 agacgtacac tgctttaact gacacgcaga cattcagcgg ctccgccgcc aatgcaaaac
     1141 tcaaccctac cccaggcagc aatgctatct caggtttgcg ggtcctttag acttgtgcaa
     1201 atatgaaaag tacatgacga ctacctgtat ttaagaatgg tgtgaggaaa tctaaccact
     1261 tattcaatgt gcagctattg ctagttgatg aatttggaat ttcagtcact cttaggaata
     1321 tgaaaccaca gaacaaatgt ccttttaaca gagatgaaat aatagtgaga attgtgagac
     1381 tacagcaaaa atctgctcag gaggaaagag ttattcagaa tagtctagga tccatagtcg
     1441 accaccatgg tggcttagat ccgggcatgt ggagacagag aagactgttg agtttgtgat
     1501 aagcactgac tctctctgcc tattggtcta ttttccctcc ctcagtggcg gtcgaggaca
     1561 aactctacgc ggacttgcca gtacgcttcg tgagcaaggg cgaggagctg ttcaccgggg
     1621 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1681 gcgagggcga gggcgacgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1741 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1801 ttagccgcta ccccgaccac ctgaagcagc acgacttctt caagagtgca atgcccgaag
     1861 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1921 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1981 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2041 atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2101 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2161 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2221 ccaacgagaa gcgcgatcat atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2281 tcggcaccga cgagctgtac aagtaaccgg tcgccaccat ggtgagcaag ggcgaggaga
     2341 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
     2401 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2461 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctaaccc
     2521 ccaacttcac ctacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2581 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2641 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2701 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     2761 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga
     2821 tgaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2881 ccaagaagcc cgtgcagctg cccggcgcct acatcgtcgg catcaagttg gacatcacct
     2941 cccacaacga ggactacacc atcgtggaac tgtacgaacg cgccgagggc cgccactcca
     3001 ccggcggcat ggacgagctg tataagtaag cggccgcgac tctagatcat aatcagccat
     3061 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3121 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3241 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     3301 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3361 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3421 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3481 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3541 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3601 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3661 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3721 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3781 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3841 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3901 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3961 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4081 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4141 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4201 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4261 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4321 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4381 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4441 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4501 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4561 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4621 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4681 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4741 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4801 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4861 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4921 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4981 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5041 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5101 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5161 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5221 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5281 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5341 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5401 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5461 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6361 cat
//
