LOCUS       sequence-177286-        4454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express shRNA against human Nup35.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4454)
  AUTHORS   Tie HC, Madugula V, Lu L
  TITLE     The development of a single molecule fluorescence standard and its 
            application in estimating the stoichiometry of the nuclear pore 
            complex.
  JOURNAL   Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi: 
            10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6.
  PUBMED    27613095
REFERENCE   2  (bases 1 to 4454)
  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: 
            "10.1016/j.bbrc.2016.09.005"; journalName: "Biochem Biophys Res 
            Commun"; date: "2016-09-30- 30"; volume: "478"; issue: "4"; pages: 
            "1694-9"
COMMENT     Sequence Label: pSUPER-retro-puro-shNup35-EcoRI
FEATURES             Location/Qualifiers
     source          1..4454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(37..636)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(120..140)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     617..636
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(656..1159)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(704..723)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1101..1124
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1170..1384
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     1305..1324
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        complement(1477..1495)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1516..1532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1516..1532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1529..1551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1540..1556
                     /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(1564..1570,1571..1588,1589..1594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1609..1630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1747..1764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1918..2506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1998..2017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2677..3468,3469..3537))
                     /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     3300..3319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3538..3642)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3668..4123)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3805..3826)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4017..4036
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4250..4272
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4264..4281
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4265..4281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4288..4307
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4288..4306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4339..4355
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4339..4355
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 cctttcggtc gggcgctgcg ggtcgtgggg cgggcgtcag gcaccgggct tgcgggtcat
       61 gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg
      121 ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg
      181 ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc
      241 gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag
      301 gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg
      361 gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac
      421 cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa
      481 gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt
      541 ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg
      601 ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgcta gcttgggctg caggtcgaaa
      661 ggcccggaga tgaggaagag gagaacagcg cggcagacgt gcgcttttga agcgtgcaga
      721 atgccgggcc tccggaggac cttcgggcgc ccgccccgcc cctgagcccg cccctgagcc
      781 cgcccccgga cccacccctt cccagcctct gagcccagaa agcgaaggag caaagctgct
      841 attggccgct gccccaaagg cctacccgct tccattgctc agcggtgctg tccatctgca
      901 cgagactagt gagacgtgct acttccattt gtcacgtcct gcacgacgcg agctgcgggg
      961 cgggggggaa cttcctgact aggggaggag tagaaggtgg cgcgaagggg ccaccaaaga
     1021 acggagccgg ttggcgccta ccggccggtg gatgtggaat gtgtgcgagg ccagaggcca
     1081 cttgtgtagc gccaagtgcc cagcggggct gctaaagcgc atgctccaga ctgccttggg
     1141 aaaagcgcct cccctacccg gtagaattcg aacgctgacg tcatcaaccc gctccaagga
     1201 atcgcgggcc cagtgtcact aggcgggaac acccagcgcg cgtgcgccct ggcaggaaga
     1261 tggctgtgag ggacagggga gtggcgccct gcaatatttg catgtcgcta tgtgttctgg
     1321 gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt ctgtatgaga
     1381 ccacagatcc ccgtacatgt ttggctggta gttcaagaga ctaccagcca aacatgtact
     1441 ttttctcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt taatttcgag
     1501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3661 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3721 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3781 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3841 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3901 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3961 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4021 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4081 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     4141 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     4201 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     4261 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagctc
     4321 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggttgccaaa cctacaggtg
     4381 gggtctttca ttcccccctt tttctggaga ctaaataaaa tcttttattt tatcgatgca
     4441 tggggtcgtg cgct
//
