LOCUS       40924_40472        8438 bp DNA     circular SYN 18-DEC-2018
DEFINITION  ShRNA expression vector pSiRPG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  AUTHORS   Storvold GL, Gjernes E, Askautrud HA, Borresen-Dale AL, Perou CM, 
            Frengen E.
  TITLE     A retroviral vector for siRNA expression in mammalian cells
  JOURNAL   Mol. Biotechnol. 35 (3), 275-282 (2007)
  PUBMED    17652791
REFERENCE   2  (bases 1 to 8438)
  AUTHORS   Stoervold GL.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2006) Institute for Medical Genetics, University 
            of Oslo, Norway, PO Box 1036 Blindern, Oslo N-0315, Norway
REFERENCE   3  (bases 1 to 8438)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8438)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Biotechnol."; date: "2007"; volume: "35"; issue: "3"; pages: 
            "275-282"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAR-2006) Institute for Medical Genetics, University of Oslo, 
            Norway, PO Box 1036 Blindern, Oslo N-0315, Norway"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    211..229
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        450..829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        830..1033
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1078..1096
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1218..1234)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1253..1669)
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    complement(1736..2077)
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     LTR             complement(2141..2657)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     promoter        3485..3589
                     /label=AmpR promoter
     CDS             3590..4447
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4621..5209
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5497..5518
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5533..5563
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5571..5587
                     /label=lac operator
                     /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)."
     repeat_region   5750..6103
                     /label=3' LTR
                     /note="3' LTR"
     CDS             complement(6176..6772)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(6782..7111)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(7188..7904)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(7939..8438)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ggtagaattc gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac
       61 taggcgggaa cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg
      121 agtggcgccc tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga
      181 aatgtctttg gatttgggaa tcttataagt tccctatcag tgatagagat ctcccgatcc
      241 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
      301 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
      361 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
      421 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
      481 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      541 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      601 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      661 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      721 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      781 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      841 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      901 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      961 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1021 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     1081 tacgactcac tatagggaga cccaagctgg ctagcgttta aacgggccct ctagactcga
     1141 gcggccgcca ctgtgctgga tatctgcaga attccaccac actggactag tggatccgag
     1201 ctcggtacca agcttatcga taccgtcgac ctcgagagat cgatctaatt ccggcgccta
     1261 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa
     1321 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct
     1381 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt
     1441 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc
     1501 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga
     1561 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat
     1621 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag
     1681 acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca
     1741 gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact
     1801 gttttaggtt ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg
     1861 actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag
     1921 ggttgcggcc gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct
     1981 agttagctaa ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa
     2041 agacagagac agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg
     2101 gtccctgggc aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg
     2161 ggcagtcaat caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca
     2221 aacagcaaga ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact
     2281 ggcgcgccga gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga
     2341 acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc
     2401 cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat
     2461 ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct
     2521 gtttggccca tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc
     2581 agttatgtat tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac
     2641 aggtggggtc tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct
     2701 atggctcgta ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga
     2761 ccactgatat cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg
     2821 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     2881 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     2941 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc
     3001 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3061 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3121 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     3181 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3241 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3361 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3421 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3601 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3661 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3721 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3781 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3841 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3901 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3961 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4021 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4081 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4141 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4201 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4261 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4321 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4381 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4441 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4501 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4741 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5221 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5281 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5341 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5401 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5461 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5521 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5581 ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa ctatttcagg
     5641 agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac ttaatattcg
     5701 actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat gaaagacccc
     5761 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
     5821 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg cagtctatcg
     5881 gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     5941 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     6001 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagcttgcca aacctacagg
     6061 tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat tttatcgatg
     6121 catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc gtggggcggg cgtcaggcac
     6181 cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg tcggcggtga
     6241 cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc tccaggaagg
     6301 cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg ctgcccagac
     6361 ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg
     6421 gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg gaaccgctca
     6481 actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg
     6541 tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc
     6601 cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccggat
     6661 cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc
     6721 tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc atggtaagct
     6781 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6841 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6901 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6961 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     7021 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     7081 ctggggactt tccacaccct aactgacaca cattccacag gtcgggtggc tctagcctta
     7141 agttcgagac tgttgtgtca gaagaatcaa gctgatctga gtccggactt gtacagctcg
     7201 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     7261 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     7321 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     7381 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     7441 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     7501 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     7561 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     7621 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     7681 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     7741 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     7801 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     7861 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtaag
     7921 ctagcttggg ctgcaggtcg aaaggcccgg agatgaggaa gaggagaaca gcgcggcaga
     7981 cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg cgcccgcccc
     8041 gcccctgagc ccgcccctga gcccgccccc ggacccaccc cttcccagcc tctgagccca
     8101 gaaagcgaag gagcaaagct gctattggcc gctgccccaa aggcctaccc gcttccattg
     8161 ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca tttgtcacgt
     8221 cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag gagtagaagg
     8281 tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg atgtggaatg
     8341 tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg ctaaagcgca
     8401 tgctccagac tgccttggga aaagcgcctc ccctaccc
//
