LOCUS       pGFP-V-RS.        7586 bp DNA     circular SYN 01-JAN-1980
DEFINITION  HuSH-29 retroviral vector with kanamycin and puromycin resistance 
            markers and a TurboGFP gene, for expressing an shRNA from the U6 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pGFP-V-RS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7586)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7586)
  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..7586
                     /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(3688..4383)
                     /label=TurboGFP
                     /note="improved green fluorescent protein from Pontellina 
                     plumata (Evdokimov et al., 2006)"
     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(5292..6098)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     LTR             6232..6701
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    6766..6965
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             7158..7574
                     /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 tcgctctttc ttcaccggca tctgcatccg gggtcttgaa
     3721 ggcgtgctgg tactccacga tgcccagctc ggtgttgctg tgatcctcct ccacgcggcg
     3781 gaaggcgaac atggggcccc cgttctgcag gatgctgggg tggatggcgc tcttgaagtg
     3841 catgtggctg tccaccacgg agctgtagta gccgccgtcg cgcaggctga aggtgcgggt
     3901 gaagctgcca tccagatcgt tatcgcccat ggggtgcagg tgctccacgg tggcgttgct
     3961 gcggatgatc ttgtcggtga agatcacgct gtcctcgggg aagccggtgc ccatcacctt
     4021 gaagtcgccg atcacgcggc cggcctcgta gcggtagctg aagctcacgt gcagcacgcc
     4081 gccgtcctcg tacttctcga tgcgggtgtt ggtgtagccg ccgttgttga tggcgtgcag
     4141 gaaggggttc tcgtagccgc tggggtaggt gccgaagtgg tagaagccgt agcccatcac
     4201 gtggctcagc aggtaggggc tgaaggtcag ggcgcctttg gtgctcttca tcttgttggt
     4261 catgcggccc tgctcggggg tgccctctcc gccgcccacc agctcgaact ccacgccgtt
     4321 cagggtgccg gtgatgcggc actcgatctc catggcgggc aggccgctct cgtcgctctc
     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 actcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc
     5281 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5341 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5401 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5461 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5521 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     5581 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5641 cgtgattgcg cctgagcgag acgaaatact cgatcgctgt taaaaggaca attacaaaca
     5701 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     5761 tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac
     5821 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     5881 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     5941 ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat
     6001 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     6061 aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     6121 atgtaagcag acagttttat tgttcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg
     6241 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     6301 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     6361 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     6421 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     6481 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     6541 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     6601 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     6661 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     6721 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
     6781 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
     6841 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
     6901 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
     6961 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     7021 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     7081 ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     7141 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttagatcact
     7201 ggaaagatgt cgagcggctc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     7261 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     7321 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     7381 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     7441 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcttccatcc gcgccgtctc
     7501 tcccccttga acctcctctt tcgaccccgc ctcaatcctc cctttatcca gccctcactc
     7561 cttctctagg cgccggccgg atcggt
//
