LOCUS       sequence-316554-        6239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged NPM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6239)
  AUTHORS   Bertwistle D, Sugimoto M, Sherr CJ
  TITLE     Physical and functional interactions of the Arf tumor suppressor 
            protein with nucleophosmin/B23.
  JOURNAL   Mol Cell Biol. 2004 Feb . 24(3):985-96.
  PUBMED    14729947
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"; journalName: "Mol Cell 
            Biol. 2004 Feb . 24(3):985-96."
COMMENT     Sequence Label: pCDNA3-FLAG-human NPM1 wt
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..79)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     785..805
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     879..898
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        879..897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             938..961
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1880..1898)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1881..1898)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1918..1935)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1924..2148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2194..2622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2281..2300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2491..2512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2631..2651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2675..2885
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2697..2893
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2744..2879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2806..2825
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2960..3754
                     /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(3014..3033)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3624..3643
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3928..4049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3965..3984)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4019..4038
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4098..4114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4098..4114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4111..4133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4122..4138
                     /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(4146..4152,4153..4170,4171..4176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4191..4212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4329..4346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4500..5088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4580..4599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5259..6050,6051..6119))
                     /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     5882..5901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6120..6224)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
       61 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      121 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      181 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      241 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      841 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      901 acccaagctt ggtaccgagc tcggatccac cgccatggac tacaaggacg atgatgacaa
      961 gggcgaattc gaagattcga tggacatgga catgagcccc ctgaggcccc agaactatct
     1021 tttcggttgt gaactaaagg ccgacaaaga ttatcacttt aaggtggata atgatgaaaa
     1081 tgagcaccag ttatctttaa gaacggtcag tttaggggct ggtgcaaagg atgagttgca
     1141 cattgttgaa gcagaggcaa tgaattacga aggcagtcca attaaagtaa cactggcaac
     1201 tttgaaaatg tctgtacagc caacggtttc ccttgggggc tttgaaataa caccaccagt
     1261 ggtcttaagg ttgaagtgtg gttcagggcc agtgcatatt agtggacagc acttagtagc
     1321 tgtggaggaa gatgcagagt cagaagatga agaggaggag gatgtgaaac tcttaagtat
     1381 atctggaaag cggtctgccc ctggaggtgg tagcaaggtt ccacagaaaa aagtaaaact
     1441 tgctgctgat gaagatgatg acgatgatga tgaagaggat gatgatgaag atgatgatga
     1501 tgatgatttt gatgatgagg aagctgaaga aaaagcgcca gtgaagaaat ctatacgaga
     1561 tactccagcc aaaaatgcac aaaagtcaaa tcagaatgga aaagactcaa aaccatcatc
     1621 aacaccaaga tcaaaaggac aagaatcctt caagaaacag gaaaaaactc ctaaaacacc
     1681 aaaaggacct agttctgtag aagacattaa agcaaaaatg caagcaagta tagaaaaagg
     1741 tggttctctt cccaaagtgg aagccaaatt catcaattat gtgaagaatt gcttccggat
     1801 gactgaccaa gaggctattc aagatctctg gcagtggagg aagtctcttt aactcgagca
     1861 tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     1921 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1981 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2041 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2101 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2161 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2221 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2281 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2341 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2401 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2461 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2521 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2581 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2821 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2881 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     2941 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3001 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3061 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3121 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3181 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3241 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3301 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3361 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3421 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3481 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3541 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3601 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3661 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3721 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3781 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3841 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3901 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     3961 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4021 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaa
//
