LOCUS       pRS.        5432 bp DNA     circular SYN 01-JAN-1980
DEFINITION  HuSH-29 retroviral vector with ampicillin and puromycin resistance 
            markers, for expressing an shRNA from the U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pRS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5432)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5432)
  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..5432
                     /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..1269
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             1351..1944
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(2316..2904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3085..3942)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3943..4047)
                     /label=AmpR promoter
     LTR             4078..4547
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    4612..4811
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             5004..5420
                     /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 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
      721 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
      781 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
      841 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
      901 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
      961 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     1021 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     1081 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     1141 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     1201 accgtcaccg ccgacgtcga gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag
     1261 cccggtgcct gagtttgttt gaatgaggct tcagtacttt acagaatcga taaaataaaa
     1321 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     1381 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     1441 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     1501 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     1561 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     1621 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     1681 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     1741 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     1801 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     1861 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     1921 acccgtcagc gggggtcttt catttccgac ttgtggtctc gctgccttgg gagggtctcc
     1981 tctgagtgat tgactacccg tcagcggggg tcttcacatg cagcatgtat caaaattaat
     2041 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     2101 acgcgcgggg agaggcggtt tgcgtattgg cgctcttccg cttcctcgct cactgactcg
     2161 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2221 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2281 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2341 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2401 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2461 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2521 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2581 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2641 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2701 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2761 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2821 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2881 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2941 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3001 acctagatcc ttttaaatta aaaatgaagt ttgcggccgc aaatcaatct aaagtatata
     3061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     3721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     4081 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4141 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4201 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     4261 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     4321 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     4381 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     4441 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     4501 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     4561 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
     4621 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     4681 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     4741 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
     4801 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     4861 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     4921 ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     4981 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     5041 atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc aagaagagac
     5101 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     5161 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     5221 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     5281 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt ccatccgcgc
     5341 cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt tatccagccc
     5401 tcactccttc tctaggcgcc ggccggatcg gt
//
