LOCUS       sequence-180074-        4454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express shRNA against human Nup88.
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-shNup88-XbaI
FEATURES             Location/Qualifiers
     source          1..4454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(48..647)
                     /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(131..151)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     628..647
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(667..1170)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(715..734)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1112..1135
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1181..1395
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     1316..1335
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        complement(1488..1506)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1527..1543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1527..1543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1540..1562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1551..1567
                     /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(1575..1581,1582..1599,1600..1605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1620..1641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1758..1775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1929..2517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2009..2028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2688..3479,3480..3548))
                     /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     3311..3330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3549..3653)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3679..4134)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3816..3837)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4028..4047
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4261..4283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4275..4292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4276..4292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4299..4318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4299..4317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4350..4366
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4350..4366
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ggtcgtgcgc tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc
       61 ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg
      121 aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc
      181 accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg
      241 ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg
      301 tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg
      361 gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc
      421 cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg
      481 cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg
      541 gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg
      601 acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatgct agcttgggct
      661 gcaggtcgaa aggcccggag atgaggaaga ggagaacagc gcggcagacg tgcgcttttg
      721 aagcgtgcag aatgccgggc ctccggagga ccttcgggcg cccgccccgc ccctgagccc
      781 gcccctgagc ccgcccccgg acccacccct tcccagcctc tgagcccaga aagcgaagga
      841 gcaaagctgc tattggccgc tgccccaaag gcctacccgc ttccattgct cagcggtgct
      901 gtccatctgc acgagactag tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc
      961 gagctgcggg gcggggggga acttcctgac taggggagga gtagaaggtg gcgcgaaggg
     1021 gccaccaaag aacggagccg gttggcgcct accggccggt ggatgtggaa tgtgtgcgag
     1081 gccagaggcc acttgtgtag cgccaagtgc ccagcggggc tgctaaagcg catgctccag
     1141 actgccttgg gaaaagcgcc tcccctaccc ggtagaattc gaacgctgac gtcatcaacc
     1201 cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa cacccagcgc gcgtgcgccc
     1261 tggcaggaag atggctgtga gggacagggg agtggcgccc tgcaatattt gcatgtcgct
     1321 atgtgttctg ggaaatcacc ataaacgtga aatgtctttg gatttgggaa tcttataagt
     1381 tctgtatgag accacagatc ccccaatcat gtaaacttct gattcaagag atcagaagtt
     1441 tacatgattg tttttctcga gggggggccc ggtaccagct tttgttccct ttagtgaggg
     1501 ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1561 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1621 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1681 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1741 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1801 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1861 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1921 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1981 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2041 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2101 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2161 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2221 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2281 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2341 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2401 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2461 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2521 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2581 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2641 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2701 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2761 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2821 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2881 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2941 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3001 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3061 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3121 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3181 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3241 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3301 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3361 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3421 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3481 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3541 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3601 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3661 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3721 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3781 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3841 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3901 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3961 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4021 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4081 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4141 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4201 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4261 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     4321 aattggagct ccaccgcggt ggcggccgct ctagaactag tggatccccc gggttgccaa
     4381 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
     4441 ttatcgatgc atgg
//
