LOCUS       pRFP-C-RS.        7454 bp DNA     circular SYN 01-JAN-1980
DEFINITION  HuSH-29 retroviral vector with chloramphenicol and puromycin 
            resistance markers and a TurboRFP gene, for expressing an shRNA from
            the U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pRFP-C-RS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7454)
  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..7454
                     /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"
     polyA_signal    complement(3091..3567)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3578..3594
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3691..4383)
                     /label=TurboRFP
                     /note="red fluorescent protein from Entacmaea quadricolor"
     promoter        complement(4427..4445)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4471..4674)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(4675..5054)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5269..5925)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     LTR             6100..6569
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    6634..6833
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             7026..7442
                     /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 gagaggacag tgccaagcaa
     3001 gcaactcaaa tgtcccaccg gttgggcatg gccaggtagc ctatgctgtg tctggacgtc
     3061 ctcctgctgg tatagttatt ttaaaatcag aaggacaggg aagggagcag tggttcacgc
     3121 ctgtaatccc agcaatttgg gaggccaagg tgggtagatc acctgagatt aggagttgga
     3181 gaccagcctg gccaatatgg tgaaaccccg tctctaccaa aaaaacaaaa attagctgag
     3241 cctggtcatg catgcctgga atcccaacaa ctcgggaggc tgaggcagga gaatcgcttg
     3301 aacccaggag gcggagattg cagtgagcca agattgtgcc actgcactcc agcttggttc
     3361 ccaatagacc ccgcaggccc tacaggttgt cttcccaact tgccccttgc tccataccac
     3421 ccccctccac cccataatat tatagaagga cacctagtca gacaaaatga tgcaacttaa
     3481 ttttattagg acaaggctgg tgggcactgg agtggcaact tccagggcca ggagaggcac
     3541 tggggagggg tcacagggat gccacccggg atctgttcag gaaacagcta tgaccgcggc
     3601 cggccgttta aacctcgagt gcggccgctt aacgcgtcgg tccgctagct gcggccgctt
     3661 cgaacgttgc gatcgctaca tcgctcttca tctgtgcccc agtttgctag ggaggtcgca
     3721 gtacttggcc acagccatct cgtgctgctc gacgtaggtc tctttgtcgg cctccttgat
     3781 tctttccagt ctgtggtcca cgaagtggaa gccgggcatc ttgaggttct tagcgggttt
     3841 cttggatctg tatgtggtct tgaaggagca gtgcaggtag cccccgccca cgagcttcag
     3901 ggccatctgg ctgtggcctc tcaggccgcc gtcagcgggg tacagcatct cggtgttggc
     3961 ctcccagccg cgtgttttct tctgcatcac agggccgttg gatgggaagt tcaccccgtt
     4021 gatcttgacg ttgtagatga tgcagccgtt ctggaagctg gtgtcctggg tagcggtcag
     4081 cacgcccccg tcttcgtatg tggtgattct ctcccatgtg aagccctcag ggaaggactg
     4141 cttaaagaag tcggggatgc cctgggtgtg gttgatgaag gctttgctgc cgtacatgaa
     4201 gctggtagcc aggatgtcga aggcgaaggg gagagggccg ccctcgacca ccttgatctt
     4261 catggtctgg gtgccctcgt agggcttgcc ttcgccctcg gatgtgcact tgaagtggtg
     4321 gttgttcacg gtgccctcca tgtacagctt catgtgcatg ttctccttga tcagctcgct
     4381 catggtggca gatctcctcg gattcagtcg acgaactccc ggccgcccta tagtgagtcg
     4441 tattacaaaa ttctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac
     4501 acgcctaccg cccatttgcg tcaacggggc ggggttatta cgacattttg gaaagtcccg
     4561 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     4621 ccgtgagtca aaccgctatc cacgcccatt ggtgtactgc caaaaccgca tcaccatggt
     4681 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccgta aggtcatgta
     4741 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcggacttgg
     4801 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4861 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4921 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     4981 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
     5041 caataatcaa tgtcaacatg gcggtcatat tggacatgag ccaatataaa tgtacatatt
     5101 atgatataga tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc
     5161 aatgactaat atggctaatt gccaatattg attcaatgta tagatcagct tggcactggc
     5221 caattggtta acaattgtag ccgcgttcta acgtcgacgt accaattacg ccccgccctg
     5281 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     5341 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     5401 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     5461 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     5521 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     5581 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     5641 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     5701 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     5761 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     5821 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     5881 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     5941 tcctgaaaat ctcgataact cattaaagct agcggccgct agcttatcga tgataagctg
     6001 tcaaacatga gtcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6061 ggttccgcgc acatttcccc gaaaagtgcc acctgcagcc tgaatatggg ccaaacagga
     6121 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     6181 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     6241 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     6301 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     6361 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     6421 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     6481 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     6541 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     6601 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt atctgtgtct
     6661 gtccgattgt ctagtgtcta tgactgattt tatgcgcctg cgtcggtact agttagctaa
     6721 ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa
     6781 ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga
     6841 gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa
     6901 acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg
     6961 tctgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     7021 attagggcca gactgttacc actcccttaa gtttgacctt agatcactgg aaagatgtcg
     7081 agcggctcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     7141 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     7201 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     7261 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     7321 ttgtacaccc taagcctccg cctcctcttc ttccatccgc gccgtctctc ccccttgaac
     7381 ctcctctttc gaccccgcct caatcctccc tttatccagc cctcactcct tctctaggcg
     7441 ccggccggat cggt
//
