LOCUS       sequence-313971-        9213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NUP188 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9213)
  AUTHORS   Itoh G, Sugino S, Ikeda M, Mizuguchi M, Kanno S, Amin MA, Iemura K, 
            Yasui A, Hirota T, Tanaka K
  TITLE     Nucleoporin Nup188 is required for chromosome alignment in mitosis.
  JOURNAL   Cancer Sci. 2013 Jul;104(7):871-9. doi: 10.1111/cas.12159. Epub 2013
            May 2.
  PUBMED    23551833
REFERENCE   2  (bases 1 to 9213)
  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: "10.1111/cas.12159";
            journalName: "Cancer Sci"; date: "2013-07"; volume: "104"; issue: 
            "7"; pages: "871-9"
COMMENT     Sequence Label: pEGFP-NUP188
FEATURES             Location/Qualifiers
     source          1..9213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(3805..4515,4516..4518,4519..4521))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3805..4515,4516..4518,4519..4521))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3847..3868)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4196..4215
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4455..4476
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4518..4527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4566..4769)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4595..4615)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4770..5073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5960..6028,6029..6820)
                     /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     complement(6178..6197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6999..7454
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7086..7105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7296..7317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7461..7582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7472..7491)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7526..7545
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gtggcgggtc tggtcgaaga gtgcaatcac ttcctcttgg agggtctcac agacattcag
       61 caccagctgg gagtactggg ggatgtcact tactttcatc tccttcattt gcaacactgt
      121 gatgaatgct gagaacacct tggccttggt cttctccatg agttgctggt cggcctgcag
      181 cactccctcc aggatctccg tcaagggtcc aaggatttca tccacagaac ctaactctct
      241 cttccactgc cggaggagga taagcagcag agtgcacttc atggagccaa ggcgaaggca
      301 gttcactgag gctggaacta ggagtaatgc tttggttcca tcaagcacgt caagaaagag
      361 ctggcgacgc accacagagt cagtcaggtg cattatatct gcatgagtgg tggcaatgat
      421 gagaagcatc ctccaggcgg acaccagcat ctggtactct aacaaggagg tgcagctgct
      481 gccttctgtt tcggccacgt gaactgccaa tgacttgaca taccctgacc agtaggcaaa
      541 gcgtttctcg atggaaaatt tcttcagtgt atcctttaat gactggtcta atgaaccctt
      601 tactacatag tatatctcca agcaaattat cttcatgatt agggcacagg tttccaggat
      661 gctgggctct gatgtttcag agggaggaga aagggttcca aacagcggac tggttaaatt
      721 ttcccaaaac ttgggtttgg ttcggaggac cagcatggca ctgtccctcc gatcctgcca
      781 cagagcatgc aaaaaggcaa tggcggcacg atgcagcagg ggtgggcacc agtatcgatc
      841 ttgctgttgg gaatcaatca gctccagcac tgcatggaga cagctccaca tcccaaggct
      901 gaattccttt gagccatcac tgccatcctt aacttccagg ttcagaaaca gttcgatgag
      961 gcctggctgg gtctctactg caacagtgag gaactctaga atcatgactt tgatgcgcat
     1021 gtcctcaatt ttgctctgca atcgggtcag gaaggcatca cgaatggcag ccgcatcatt
     1081 gcccagacaa gcatacactg acattggggc caccgtggcc agacgtttca gcagctgaat
     1141 ggcaagacgt ggcaaagcag ggtcatgttt gtggtagatg tatttggcta gaacagcaat
     1201 gaggttgttt ccatgagcac catgttgtga gagagcctgt tccagggggg acaccacatt
     1261 agaaggaggt ttcagccgaa taacattgtt ggtgacggag aatgccagtt tcactgtctt
     1321 gatcagcagc tggccctgcc cctggccctc tgccccatca ctagggtgcc ttcgaggctg
     1381 agcagccatc accatgtcaa tggtgtccac gccaatgccc atgatattga taactgtctg
     1441 tcctgcttct gtgtatgcca ggctgcagat gcagagaaac tgcaggctgg gagtatgact
     1501 gctgtgcagg tctgtctcgt ggcacaggtt cagtatcgca tgaatcagct ccaagatcag
     1561 gcaaccaatc tgttccctca ctccatgaga gttgtagcgc cacttatggt agctgggaag
     1621 catctccttc agcacaaaca ttacacaggg tacaagtcct tggctctggg tactaccaag
     1681 ttgccccttg acaagggtgg tgatcaagcg cagaaaggca atagtaaccc catactcgcc
     1741 ctgaggctgt tcactgttca tcaagaggtt tccgtaccct ccagcattca tcccttccgc
     1801 actaatcatc tgactcaggc tggagacagg atgggccaca aatggtaaaa aacctgtgtg
     1861 acgaagatca gtccagacct ttgctggatt gcgggcagcc aaaacagtta agcagttgac
     1921 acaagaagca atgacatcca caggtgggga gatcactgtc gttaaccgct gcagcagcat
     1981 gtagatgcga gatgtgatgg gcaggagaca gtctgctatc gacaggtctg tactgatgac
     2041 cttatggacg agatcaatga tgggtttgac tcgctggcag tgctgaatca catctgcagt
     2101 tgaaacaaca tgaagcaaca tttcaatctc gcaggtaaag agggtccagc tgctatagga
     2161 gtattcccag cgtaccaggt atgccctatc atccaacatt acttggccca cagtgccttg
     2221 aggtatgcga aggttggttt gacccccaag gggataaagg agtttgggtg tttgtctccg
     2281 ccaaagagtt ccatcttcat gggagatcac atcatgaggc ttgtgtttat aaagttcatt
     2341 gtagaaagac atcttatcca agaagctata cacctttttg gctgtggact tccctgatac
     2401 cagggctcgg agcagttgca ggagtgggga gagaaggtgg ggaaacattc cacacacact
     2461 gtccagaatg atcccaaggc cagaagttgg ctctgttccc cagaacagtt ccggaagaga
     2521 agggtcggcc aatacttcac atgctgtatc aattatatcc tgctgattgc ccagggtgtg
     2581 cagctccaac gaggtcagaa cgaaagagag cagtccatag acacacatgc atgcagtgct
     2641 ggtggtgcaa tcatttcccc cactggcaag ggactggagc aatcgggtca agtactgaaa
     2701 cacattcagc tggatggctg tgccacctat cttccggacc acactgcttg tctcttctgg
     2761 gttcagagtg tgacggagga gagcccaggc caaaagcact ggggcatgat gtggaatgtc
     2821 cccaaaggtc aacattaaac agtccatatc ctgacaaata agcccatcct gcgcaaactg
     2881 atgcagttct cttctgtcat ccaaagcaca cttatgcaag gactcgatat ccatgccctc
     2941 caccaggatg agggcactga agtagccaat ccgatctaca aaaggatcca tagtctcatc
     3001 caccaggtgc ctattggtct gcctactacc aaatccttgc tctttaaaca tcttggttaa
     3061 tacaagtaag tcactgggtg ccatctcaaa gtatgcataa taaaggaaaa taatttctag
     3121 cagcatggac tgttcccgaa ggcactgaac aaaccagcga gacacttggc gctctgtcat
     3181 gagatttcca tgtgtctccc aagttggtgc ttcagtttta taaagctctt cgaactgctg
     3241 tctgtatttt gaaactagtt ccttctccaa tttatcaaca cagtctgcat attcaaccct
     3301 ataggggtgt ctttcatctt ggaagtaagt gagaaggtgt aagacacaac gaagaataca
     3361 ggttctttct tcataataat aatctgcaat cttcaggatt aaggcctggc tctgcctctc
     3421 atcttgcagt actgtcttta ctgagtcccg agtacccctg tagtcctctt gcaggtaaca
     3481 ctggagtaac tgcacactct gttcttcatc aagacccaaa aacttgctga ttcttaaacc
     3541 cagttccttc aatggtgaag ctacatcttt attagctttc actttttcag ctgaacttgg
     3601 actgggaggt ttgtagtaag aaagcccctc taacaatcgc cgccaatgtt tattcagttc
     3661 tgcctcaatc tgactcagct ctctcagagc tgaccttcca agcagaatag tccacagttc
     3721 tctactgctc ctcacacacg gcccgccggc ggcctcgagg accatggtgt cgactgcaga
     3781 attcgaagct gatctgagtc cggacttgta cagctcgtcc atgccgagag tgatcccggc
     3841 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     3901 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     3961 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     4021 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     4081 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     4141 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     4201 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     4261 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     4321 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     4381 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     4441 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     4501 ctcctcgccc ttgctcacca tggtggcgac cggtagcgct agcggatctg acggttcact
     4561 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     4621 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     4681 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     4741 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     4801 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     4861 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     4921 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     4981 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     5041 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     5101 aatgaccccg taattgatta ctattaataa ctaatgcatg gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagacaataa ccctgataaa tgcttcaata
     5941 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6001 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6061 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6121 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6181 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6241 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6301 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6361 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6421 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6481 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6541 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6601 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6661 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6721 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6781 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6841 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6901 cctttttgat aatctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     7021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7441 aacgcttaca atttacgcgt taagatacat tgatgagttt ggacaaacca caactagaat
     7501 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7561 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     7621 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     7681 ttatgatcag ttatctagat ccggtggatc gcggccgcat ctttgcatat gccggacaaa
     7741 cgcctgcacc aactggatca gaggctcctg actctcaggg ctggctttgg agagagaggt
     7801 ggacttgccc tgcggcgagg ggaggacacc agtggcaggg gagctggggg ctccccggcg
     7861 gaagtagcgc gagaggctgg acagcagcgt gctcaactca gagctgagct cctgcttcat
     7921 ccgctgtttg tcccgggggt gcacagccgg gtccctaagg taccgcatcg cctgagagat
     7981 gagcaggtag aagcagtttt ccatggtaaa catcaggagg gactttaacg tcctggtccc
     8041 ttctgcctgt gtgctgagcc ccactgcctg ggtgaggggc tcctttttct tgtccagctc
     8101 tccaagcatg ttgagggcca cattcactgt ggccagaagg gtcccgaagg agggggccac
     8161 ttcggagtca aaggtgggag tggtaaagct cagggcaaac aggaagttgt attcagcaag
     8221 gtccagggac tgatccagca gaatctggca gacatctggg gtgaagtggc gcagggctgc
     8281 cagcgtcttg ctgaggatct tgagaaggcc atactggact gtgtgcaagg cctgctgctc
     8341 tgatgcctct gtgtcagcag cgggctgctt tgaggaggag ggggcagcag aagcagcaga
     8401 cggtggcctc tggactcgct gggcaacagc tgaggggagg ccatccccat ttttgttctg
     8461 taagtaatgc tgcagcatct ttcgactgtg caggagagag gtacatgcct ggcacaagta
     8521 acccaggttg acctggatat cacgcatgag ctgaggcagg tggaagtgcc actccttcat
     8581 gaagttagag agctgcagaa taaaacccac ggtgtggtcc gcctcctcca ggcaggccag
     8641 actctgcact gtcctcactg cgttgaggca ctgtaaggtc cgctcctggt ggacacccac
     8701 gaagtccagg gcctcaggca ggaagttgta gcgcagagtt ttgagcagct gctccatcag
     8761 ggacatggac aggcggtaga ctcctggcca agagggggca tccagggact tccgagaggc
     8821 actaggtgtc tgtgctgtgc cgttggtgct cagctggtac acactcagaa ggggcaaaca
     8881 aatgctctgg gtgatgccag ctccagccac tgctgtggct ccctgctgag tgcgagccag
     8941 ggtgaggagc agatgcaatg tggcctcagt gaaatgcagg ttctgcttca tgcgaaggct
     9001 cacctctaga gtggtgagga gggtgggtag gatggggagc ctgcgggtta cctgcagcca
     9061 ggagtcacca tcctcgtcta cctcacacag ctccttggcc aggtgcaggc ccaggacaca
     9121 caccccatca cgctggtccc tgtgccggga ccgagaacag tcgtcagtct ccatgctgtc
     9181 cttgtcctct gtggcactgc ctaatgccag act
//
