LOCUS       sequence-264365-        6239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-NPM1wildtype (human) in mammalian cells through 
            transient transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6239)
  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 6239)
  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-NPM1wt
FEATURES             Location/Qualifiers
     source          1..6239
                     /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"
     promoter        complement(1864..1882)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1865..1882)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1902..1919)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1908..2132
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2178..2606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2265..2284)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2475..2496
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2615..2635)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2659..2869
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2681..2877
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2728..2863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2790..2809
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2944..3738
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2998..3017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3608..3627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3912..4033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3949..3968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4003..4022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4082..4098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4082..4098)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4095..4117)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4106..4122
                     /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(4130..4136,4137..4154,4155..4160))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4175..4196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4313..4330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4484..5072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4564..4583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5243..6034,6035..6103))
                     /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     5866..5885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6104..6208)
                     /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 tctggcagtg gaggaagtct ctttaactcg agcatgcatc tagagggccc
     1861 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     1921 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1981 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2041 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2101 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2161 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2221 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2281 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2341 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2401 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2461 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2521 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2581 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2641 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2701 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2761 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2821 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2881 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     2941 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3001 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3061 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3121 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3181 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3241 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3301 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3361 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3421 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3481 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3541 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3601 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3661 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3721 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3781 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3841 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3901 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3961 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4021 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4081 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4141 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4201 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4261 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4321 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4381 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4441 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4501 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4561 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4621 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4681 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4741 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4801 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4861 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4921 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4981 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5041 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5101 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5161 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5221 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5281 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5341 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5401 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5461 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5521 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5581 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5641 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5701 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5761 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5821 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5881 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5941 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6001 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6061 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6121 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6181 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
