LOCUS       pB-RS.        5231 bp DNA     circular SYN 01-JAN-1980
DEFINITION  HuSH-29 retroviral vector with ampicillin and blasticidin resistance
            markers, for expressing an shRNA from the U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pB-RS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5231)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5231)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The shRNA cassette consists of a 29-bp gene-specific sequence, 
            followed by a 7-bp loop, followed by the reverse complement of the 
            29-bp sequence, followed by a TTTTTT terminator.
FEATURES             Location/Qualifiers
     source          1..5231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    334..385
                     /label=shRNA site
                     /note="shRNA site"
                     /note="shRNA insertion site"
     promoter        441..651
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     CDS             673..1068
                     /label=BSD
                     /note="blasticidin S deaminase"
     LTR             1150..1743
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(2115..2703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2884..3741)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3742..3846)
                     /label=AmpR promoter
     LTR             3877..4346
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    4411..4610
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             4803..5219
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
ORIGIN
        1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc
       61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt
      121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
      181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
      241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
      301 tcttgggttt atatatcttg tggaaaggac gcgggatcca ctggaccagg cagcagcgtc
      361 agaagacttt tttttggaaa agcttgtcga ccctgtggaa tgtgtgtcag ttagggtgtg
      421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      481 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      541 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      601 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      661 agctagctta ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
      721 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
      781 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
      841 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
      901 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
      961 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     1021 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agtttgtttg
     1081 aatgaggctt cagtacttta cagaatcgat aaaataaaag attttattta gtctccagaa
     1141 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     1201 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
     1261 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     1321 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     1381 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
     1441 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     1501 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     1561 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg
     1621 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     1681 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     1741 atttccgact tgtggtctcg ctgccttggg agggtctcct ctgagtgatt gactacccgt
     1801 cagcgggggt cttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     1861 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1921 gcgtattggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2521 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2821 aaatgaagtt tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2881 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2941 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3001 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3061 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3121 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3181 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3241 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3301 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3361 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3421 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3481 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     3541 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3601 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3661 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3721 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3781 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3841 tccgcgcaca tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat
     3901 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     3961 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4021 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     4081 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     4141 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     4201 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     4261 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     4321 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
     4381 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
     4441 cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta
     4501 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
     4561 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
     4621 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     4681 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     4741 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     4801 agggccagac tgttaccact cccttaagtt tgaccttaga tcactggaaa gatgtcgagc
     4861 ggctcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     4921 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     4981 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     5041 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     5101 tacaccctaa gcctccgcct cctcttcttc catccgcgcc gtctctcccc cttgaacctc
     5161 ctctttcgac cccgcctcaa tcctcccttt atccagccct cactccttct ctaggcgccg
     5221 gccggatcgg t
//
