LOCUS       sequence-177293-        6351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express shRNA against human Nup133.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  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 6351)
  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-shNup133-1
FEATURES             Location/Qualifiers
     source          1..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..639)
                     /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(123..143)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     620..639
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(664..1163)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(712..731)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1105..1128
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     promoter        1174..1388
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     1309..1328
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     CDS             complement(1471..1887)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1487..1503)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1519..1541)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    1954..2295
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             2359..2875
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3075..3094)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3222..3244)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3438..3457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3557..3579
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3617..3635)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3703..3807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3808..3876,3877..4668)
                     /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(4026..4045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4839..5427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5328..5347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5581..5598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5715..5736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5751..5756,5757..5774,5775..5781)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5789..5805
                     /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)."
ORIGIN
        1 gctcctttcg gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt
       61 catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag
      121 ccgctcgtag aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc
      181 gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg
      241 gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc
      301 caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg
      361 cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc
      421 caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag
      481 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg
      541 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc
      601 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gtaagctagc ttgggctgca
      661 ggtcgaaagg cccggagatg aggaagagga gaacagcgcg gcagacgtgc gcttttgaag
      721 cgtgcagaat gccgggcctc cggaggacct tcgggcgccc gccccgcccc tgagcccgcc
      781 cctgagcccg cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca
      841 aagctgctat tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc
      901 catctgcacg agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag
      961 ctgcggggcg ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc
     1021 accaaagaac ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag
     1081 gccacttgtg tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct
     1141 tgggaaaagc gcctccccta cccggtagaa ttcgaacgct gacgtcatca acccgctcca
     1201 aggaatcgcg ggcccagtgt cactaggcgg gaacacccag cgcgcgtgcg ccctggcagg
     1261 aagatggctg tgagggacag gggagtggcg ccctgcaata tttgcatgtc gctatgtgtt
     1321 ctgggaaatc accataaacg tgaaatgtct ttggatttgg gaatcttata agttctgtat
     1381 gagaccacag atcccctgtt cagggacaaa tataattcaa gagattatat ttgtccctga
     1441 acatttttct cgagagatcg atctaattcc ggcgcctaga gaaggagtga gggctggata
     1501 aagggaggat cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg
     1561 aggaagagga ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa
     1621 agccaaggct tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg
     1681 tgaaaagacc ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg
     1741 ccatccgacg ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt
     1801 gacatctacc gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa
     1861 acttaaggga gtggtaacag tctggcccta attttcagac aaatacagaa acacagtcag
     1921 acagagacaa cacagaacga tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt
     1981 ccaaaccgaa agcaaaaatt cagacggagg cgggaactgt tttaggttct cgtctcctac
     2041 cagaaccaca tatcctgacg gggtcggatt ccacatcgac tcccttcctc aggtcgggcc
     2101 acaaaaacgg cccccaaagt ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa
     2161 ctcgtcagtt ccaccacggg tccgccagat acagagctag ttagctaact agtacagacg
     2221 caggcgcaaa cattagatgc cggcacaaac acgcacaaag acagagacag acacgaaacg
     2281 accgctggcc agcttacctc ccggcggggg ggtcggtggt ccctgggcag gggtctccaa
     2341 atctcggtgg aacctccaaa tgaaagaccc ccgaggtggg cagtcaatca atctgaggag
     2401 accctcccaa ggatcagcga gtccacgatt cggatgcaaa cagcaagagg ctttattggg
     2461 aatacgggta cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt
     2521 gggctctttt attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga
     2581 ttggttagtt caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc
     2641 tgatggttct ctagaaactg ctgagggcgg gaccgcatct ggggaccatc tgttcttggc
     2701 cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata ttctgctgtc
     2761 tctctgttcc taaccttgat ctgaacttct ctattctcag ttatgtattt tccatgcctt
     2821 gcaaaatggc gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc
     2881 ctttttctgg agactaaata aaatctttta ttttatctat ggctcgtact ctataggctt
     2941 cagctggtga tattgttgag tcaaaactag agcctggacc actgatatcc tgtctttaac
     3001 aaattggact aatcgcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     3061 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     3121 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     3181 gttgggcgcc atctccttgc atgcaccatt ccttgcgccg tcgttttaca acgtcgtgac
     3241 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3301 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3361 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3421 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3481 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3541 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3601 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3661 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3781 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3841 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3901 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3961 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4021 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4081 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4141 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4201 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4261 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4321 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4381 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4441 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4501 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4561 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4621 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4681 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4741 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4801 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4861 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4921 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4981 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5041 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5101 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5161 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5221 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5281 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5341 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5401 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5461 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5521 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5581 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5641 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5701 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5761 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca caattctccc
     5821 cagctctttc tgcgctgtcc ataatgaact atttcaggag tattggagta tgtggattca
     5881 aaacagtggc taattgagtc aggattactt aatattcgac tcctaccctc agaatgttgt
     5941 tctccaactt caagaaactg ttacccatga aagacccccg ctgacgggta gtcaatcact
     6001 cagaggagac cctcccaagg aacagcgaga ccacaagtcg gatgcaactg caagagggtt
     6061 tattggatac acgggtaccc gggcgacgca gtctatcgga ggactggcgc gccgagtgag
     6121 gggttgtggg ctcttttatt gagctcgggg agcagaagcg cgcgaacaga agcgagaagc
     6181 gaactgattg gttagttcaa ataaggcaca gggtcatttc aggtccttgg ggcaccctgg
     6241 aaacatctga tggttctcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt
     6301 tttctggaga ctaaataaaa tcttttattt tatcgatgca tggggtcgtg c
//
