LOCUS       sequence-264364-        6284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-NPM1c (cytoplasmic mutant, human) in mammalian cells 
            through transient transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6284)
  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 6284)
  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: pcDNA3.1-Flag-NPM1c
FEATURES             Location/Qualifiers
     source          1..6284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1869..1885)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1870..1886)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1909..1927)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1910..1927)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1947..1964)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1953..2177
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2223..2651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2310..2329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2520..2541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2660..2680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2704..2914
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2726..2922
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2773..2908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2835..2854
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2989..3783
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHPIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3043..3062)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3653..3672
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3957..4078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3994..4013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4048..4067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4127..4143)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4127..4143)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4140..4162)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4151..4167
                     /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)."
     promoter        complement(join(4175..4181,4182..4199,4200..4205))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4220..4241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4358..4375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4529..5117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4609..4628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5288..6079,6080..6148))
                     /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     5911..5930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6149..6253)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccaccgccat ggactacaag gacgatgatg acaagggcga attcgaagat
      961 tcgatggaca tggacatgag ccccctgagg ccccagaact atcttttcgg ttgtgaacta
     1021 aaggccgaca aagattatca ctttaaggtg gataatgatg aaaatgagca ccagttatct
     1081 ttaagaacgg tcagtttagg ggctggtgca aaggatgagt tgcacattgt tgaagcagag
     1141 gcaatgaatt acgaaggcag tccaattaaa gtaacactgg caactttgaa aatgtctgta
     1201 cagccaacgg tttcccttgg gggctttgaa ataacaccac cagtggtctt aaggttgaag
     1261 tgtggttcag ggccagtgca tattagtgga cagcacttag tagctgtgga ggaagatgca
     1321 gagtcagaag atgaagagga ggaggatgtg aaactcttaa gtatatctgg aaagcggtct
     1381 gcccctggag gtggtagcaa ggttccacag aaaaaagtaa aacttgctgc tgatgaagat
     1441 gatgacgatg atgatgaaga ggatgatgat gaagatgatg atgatgatga ttttgatgat
     1501 gaggaagctg aagaaaaagc gccagtgaag aaatctatac gagatactcc agccaaaaat
     1561 gcacaaaagt caaatcagaa tggaaaagac tcaaaaccat catcaacacc aagatcaaaa
     1621 ggacaagaat ccttcaagaa acaggaaaaa actcctaaaa caccaaaagg acctagttct
     1681 gtagaagaca ttaaagcaaa aatgcaagca agtatagaaa aaggtggttc tcttcccaaa
     1741 gtggaagcca aattcatcaa ttatgtgaag aattgcttcc ggatgactga ccaagaggct
     1801 attcaagatc tctgtctggc agtggaggaa gtctctttaa gaaaatagga attcgatatc
     1861 aagcttatcg ataccgtcga cctcgagcat gcatctagag ggccctattc tatagtgtca
     1921 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1981 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2041 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2101 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2161 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2221 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2281 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2401 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2521 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2581 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2641 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2821 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2941 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3001 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3061 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3121 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3181 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3241 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3301 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3361 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcccat cgagcgagca
     3421 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3481 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3541 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3601 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3661 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3721 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3781 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3841 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3901 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3961 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4021 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4081 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4141 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4201 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4261 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4321 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4381 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4441 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4501 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4561 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4921 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4981 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5041 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5101 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5161 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5221 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5281 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5341 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5401 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5461 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5521 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5581 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5641 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5701 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5761 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5821 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5881 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5941 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6001 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6061 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6121 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6181 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6241 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
