LOCUS       sequence-78547-e        6354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mouse phosphorylation-mimicking Npm2 (p-Npm, N533).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  TITLE     Ishii Lab histone variant plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6354)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDON-AI-Npm2(N533)
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        248..451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     402..422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    672..1029
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     promoter        1998..2208
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2020..2216
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2067..2202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2129..2148
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2283..3077
                     /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(2337..2356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2947..2966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3126..3719
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3735..3751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3735..3751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3748..3770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3759..3775
                     /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(3783..3789,3790..3807,3808..3813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3828..3849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3966..3983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4137..4725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4217..4236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4896..5687,5688..5756))
                     /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     5519..5538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5757..5861)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5929..5947
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5985..6007)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6107..6126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6320..6342
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6334..6351
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6335..6351
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttatgt gaaagacccc acctgtaggt ttggcaagct agcgggactt tccattgacg
       61 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      121 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      181 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      241 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      301 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      361 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      421 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgcgcca gtcttccgat
      481 agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
      541 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
      601 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
      661 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
      721 actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc
      781 cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggac
      841 ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt
      901 tggtgcaccc cccttagagg agggatatgt ggttctggta ggagacgaga acctaaaaca
      961 gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct
     1021 tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1081 aaatacgcgt cggcttcctt tgtccccaat ctgggcgcgc gccggcgccc cctggcggcc
     1141 taaggactcg gcgcgccgga agtggccagg gcgggggcga cttcggctca cagcgcgccc
     1201 ggctattctc gcagctcacc gctagcgttt aaacttaagc ttggtaccga gctcggatcc
     1261 gtcgacgttt cgaggtaaat cgacatgagt cgccacgatg atgacgacgt ggacgaaacc
     1321 acagcaaaaa acatgctctg gggtagtgaa ctcaatcagg aaaagcagac ttgcaccttt
     1381 agaggccaag gcgagaagaa ggacagctgt aaactcttgc tcagcacgat ctgcctgggg
     1441 gagaaagcca aagaggaggt gaaccgtgtg gaagtcctct cccaggaagg cagaaaacca
     1501 ccaatcacta ttgctacgct gaaggccgac gtcctgccca tggtcactgt gtcaggtata
     1561 gagcttgatc ctccagtaac ttttcggctc agggatggct caggacctgt gttcctcagt
     1621 ggcctggaat gttatgagac ttcggacctg acctgggaag atgacgagga agaggaggaa
     1681 gaggaggagg aagaggatga agatgaggat gcagatatat cgctagagga gatacctgtc
     1741 aaacaagtca aaagggtggc tccccagaag cagatgagca tagcaaagaa aaagaaggtg
     1801 gaaaaagaag aggatgaaac agtagtgagg cccgaccctc aggacaagga tccctggaag
     1861 aaggagaaat ctgatcccag agcaaagaag ccagtgacca agaaatgacc tcattctagc
     1921 caccatgact tcgatcgaat taacgcatgc atctcaatta gtcagcaacc aggtgtggaa
     1981 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2041 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2101 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2161 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2221 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2281 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2341 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2401 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2461 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2521 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2581 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2641 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2701 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2761 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     2821 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2881 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2941 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3001 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3061 ttgacgagtt cttctgactc gaggataaaa taaaagattt tatttagtct ccagaaaaag
     3121 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     3181 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     3241 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     3301 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     3361 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     3421 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     3481 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     3541 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
     3601 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     3661 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattg
     3721 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3781 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3841 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5341 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5521 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5581 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5641 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5701 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5941 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6001 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6061 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     6121 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6181 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6241 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6301 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgcc
//
