LOCUS       pSUPER-retro-puro-shNup133-2              6351 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  To express shRNA against human Nup133.
ACCESSION   .
VERSION     .
KEYWORDS    pSUPER-retro-puro-shNup133-2
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 Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6351
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        complement(1488..1702)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(1548..1567)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        1713..2212
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1748..1771)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2145..2164
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2237..2836
                     /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(2237..2256)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2733..2753
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    3422..3438
                     /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(3446..3476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3491..3512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3629..3646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3800..4388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3880..3899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4559..5419)
                     /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     5182..5201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5420..5524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5592..5610
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5648..5670)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5770..5789
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5983..6005
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6133..6152
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatcgatc tctcgagaaa aaggaggtga
     1441 aagacctgct atctcttgaa tagcaggtct ttcacctccg gggatctgtg gtctcataca
     1501 gaacttataa gattcccaaa tccaaagaca tttcacgttt atggtgattt cccagaacac
     1561 atagcgacat gcaaatattg cagggcgcca ctcccctgtc cctcacagcc atcttcctgc
     1621 cagggcgcac gcgcgctggg tgttcccgcc tagtgacact gggcccgcga ttccttggag
     1681 cgggttgatg acgtcagcgt tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca
     1741 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     1801 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     1861 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     1921 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     1981 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     2041 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     2101 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     2161 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     2221 cccaagctag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     2281 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     2341 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     2401 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     2461 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     2521 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     2581 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     2641 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     2701 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     2761 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     2821 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     2881 accccatgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     2941 agaccccacc tgtaggtttg gcaagctaga gaaccatcag atgtttccag ggtgccccaa
     3001 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     3061 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     3121 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     3181 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     3241 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg
     3301 gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact
     3361 ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa
     3421 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3481 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3541 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3601 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4201 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4501 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4561 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4621 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4681 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4741 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4801 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4861 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4921 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4981 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5041 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5101 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5161 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5221 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5281 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5341 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5401 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5461 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5521 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5581 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     5641 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5701 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     5761 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     5821 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     5881 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5941 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     6001 aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     6061 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     6121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     6181 gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat
     6241 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
     6301 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg g
//