LOCUS       sequence-309660-        4475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4475)
  AUTHORS   Kowalczyk SW, Kapinos L, Blosser TR, Magalhaes T, van Nies P, Lim 
            RY, Dekker C
  TITLE     Single-molecule transport across an individual biomimetic nuclear 
            pore complex.
  JOURNAL   Nat Nanotechnol. 2011 Jun 19;6(7):433-8. doi: 10.1038/nnano.2011.88.
  PUBMED    21685911
REFERENCE   2  (bases 1 to 4475)
  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/nnano.2011"; journalName: "Nat Nanotechnol"; date: 
            "2011-06-19- 19"; volume: "6"; issue: "7"; pages: "433-8"
COMMENT     Sequence Label: pPEP-TEV cNup98 (1-498)
FEATURES             Location/Qualifiers
     source          1..4475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             86..103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             224..244
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1815..1833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1829..1876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2051..2056,2057..2073,2074..2079))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2157..2174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2328..2916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2408..2427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3087..3878,3879..3947))
                     /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     3710..3729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3948..4052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4120..4138
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4176..4198)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4298..4317
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgggccacca tcaccatcat cacatgcaaa aatacttaga
      121 agataaagct caagaattgg tgcgactgga aggagaggtt cgctccctcc ttaaggacat
      181 aagtgagaaa gttgcggttt acagcaccgg cggtggtggc ggtgagaatt tatattttca
      241 agggggatcc tgttgctgtt ttaacaaatc atttggaaca ccctttgggg gtggcacagg
      301 tggctttggc acaacttcaa catttggaca gaatactggc tttggcacta ctagtggagg
      361 ggcatttgga acatctgcat ttggttctag caacaatact ggaggcctct ttggaaattc
      421 acagactaaa ccaggaggat tgtttggaac cagttcattt agccagccag ctacctccac
      481 aagcactggc tttgggtttg gtacgtcaac aggaacagca aataccttgt ttggaactgc
      541 aagcacaggg accagtctct tctcatccca aaacaatgcc tttgcacaaa ataaaccaac
      601 tggctttggc aattttggaa ccagtactag cagtggagga ctctttggaa ccacaaatac
      661 cacctctaat ccttttggca gcacatctgg ctccctcttt gggccaagta gttttacagc
      721 tgctcctact gggactacta ttaaatttaa ccctccaact ggtacagata ctatggtcaa
      781 agctggagtt agcactaaca taagtaccaa gcaccagtgt attactgcta tgaaagaata
      841 tgaaagcaag tcactagagg aacttcgttt agaggattat caggctaaca ggaagggccc
      901 acagaaccag gtgggagcag gtaccacaac tggcttgttt gggtcttctc cagccacttc
      961 cagcgcaaca ggactcttca gctcctccac cactaattca ggctttgcat atggtcagaa
     1021 caaaactgcc tttggaacta gtacaactgg atttggaaca aatccaggtg gtctctttgg
     1081 ccaacagaat cagcagacta ccagcctctt cagcaaacca tttggccagg ctacaaccac
     1141 ccagaacact ggcttttcct ttggtaatac cagcaccata ggacagccaa gcaccaacac
     1201 catgggatta tttggagtaa cccaagcctc acagcctgga ggtctttttg ggacagctac
     1261 aaacaccagc actgggacag catttggaac aggaacaggt ctctttgggc agaccaatac
     1321 tggatttggt gctgttggtt cgaccctgtt tggcaataac aagcttacta catttggaag
     1381 cagcacaacc agtgcacctt catttggtac aaccagtggc gggctctttg gttttggcac
     1441 aaataccagt gggaatagta tttttggaag taaaccagca cctgggactc ttggaactgg
     1501 gcttggtgca ggatttggaa cagctcttgg tgctggacag gcatctttgt ttgggaacaa
     1561 ccaacctaag attggagggc ctcttggtac aggagccttt ggggcccctg gatttaatac
     1621 tacgacagcc actttgggct ttggagcccc ccaggcccca gtagctttga cagatccaaa
     1681 tgcttctgct gcccagcagg ctgttctcca gcagcacatc aatagtctaa catactcacc
     1741 ttttggagac tcttaagaat tctaagatcc ggctgctaac aaagcccgaa aggaagctga
     1801 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1861 cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc
     1921 agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg
     1981 atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat
     2041 aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt actgcattaa
     2101 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2161 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3001 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3061 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3121 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3181 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3241 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3301 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3361 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3421 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3481 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3541 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3601 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3661 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3721 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3781 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3841 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3901 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3961 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4021 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4081 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4141 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4201 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4261 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4321 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4381 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4441 tcgctattac gccagtgatc tcgatcccgc gaaat
//
