LOCUS       pSUPER-retro-puro-shNup93-SpeI                4454 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expression of shRNA targeting at Nup93.
ACCESSION   .
VERSION     .
KEYWORDS    pSUPER-retro-puro-shNup93-SpeI
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 Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4454
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     276..295
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             363..962
                     /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(363..382)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     859..879
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1098..1114)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1098..1114)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1146..1165)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1147..1165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1172..1189)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1172..1188)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1181..1203)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1330..1785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1417..1436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1627..1648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1811..1915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1916..2776
                     /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(2134..2153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2947..3535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3436..3455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3689..3706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3823..3844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3859..3889
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3897..3913
                     /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)."
     primer_bind     3902..3924
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3921..3937
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3921..3937
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3958..3976
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4069..4283)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(4129..4148)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(4329..4352)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
       61 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
      121 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
      181 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
      241 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
      301 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagcta
      361 gcatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
      421 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
      481 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
      541 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
      601 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
      661 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
      721 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
      781 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
      841 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
      901 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
      961 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
     1021 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     1081 ggtttggcaa cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     1141 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     1201 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1261 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     1321 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1381 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1441 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1501 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1561 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1621 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1681 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1741 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2101 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2221 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2461 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2521 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2761 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2821 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3601 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3661 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3721 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3781 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     3841 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     3901 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat
     3961 taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgagaa aaacacaaag
     4021 cctattatca actctcttga agttgataat aggctttgtg ggggatctgt ggtctcatac
     4081 agaacttata agattcccaa atccaaagac atttcacgtt tatggtgatt tcccagaaca
     4141 catagcgaca tgcaaatatt gcagggcgcc actcccctgt ccctcacagc catcttcctg
     4201 ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac tgggcccgcg attccttgga
     4261 gcgggttgat gacgtcagcg ttcgaattct accgggtagg ggaggcgctt ttcccaaggc
     4321 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     4381 ggcctcgcac acattccaca tccaccggcc ggtaggcgcc aaccggctcc gttctttggt
     4441 ggccccttcg cgcc
//