LOCUS       V010486                 8691 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010486
VERSION     V010486
KEYWORDS    pGFP-C-shLenti
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8691)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8691)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            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..8691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        237..616
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..819
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             834..1014
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1683..1916
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2101..2145
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             2294..2335
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2443..2560
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2707..2947
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2951..3008
                     /label="shRNA site"
                     /note="shRNA site"
                     /note="shRNA insertion site"
     promoter        3064..3274
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     CDS             3296..3892
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     enhancer        4301..4680
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4682..4881
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4910..4928
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5036..5731
                     /label="TurboGFP"
                     /note="improved green fluorescent protein from Pontellina
                     plumata (Evdokimov et al., 2006)"
     protein_bind    complement(5786..5819)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5875..6463
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6466..6482)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             6992..7172
                     /label="3' LTR (truncated)"
                     /note="3' LTR (truncated)"
                     /note="truncated 3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7234..7822)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7902..8558)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(8559..8663)
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctaggaa ttccccagtg gaaagacgcg
     2641 caggcaaaac gcaccacgtg acggagcgtg accgcgcgcc gagcgcgcgc caaggtcggg
     2701 caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2761 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2821 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2881 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tgggtttata tatcttgtgg
     2941 aaaggacgcg ggatccactg gaccaggcag cagcgtcaga agactttttt ttggaacgtc
     3001 tcaagcttgt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3061 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3241 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctagc ttaccatgac
     3301 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3361 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3421 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3481 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3541 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3601 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3661 gtggttcctg gccaccgtcg gcgtgtcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3721 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3781 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3841 cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctgagtttg
     3901 tttgaatgag gcttcagtac tttacagaat cgataaaata aaagatttta tttagtctcc
     3961 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     4021 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     4081 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4141 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4201 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     4261 ctcagcagtt tctagacatg tccaatatga ccgccatgtt gacattgatt attgactagt
     4321 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4381 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4441 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4501 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4561 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4621 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4681 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
     4741 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4801 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
     4861 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
     4921 actatagggc ggccgggaat tgatccggta ccgaggagac tgccgccgcg atcgccggcg
     4981 cgccagatct caagcttaac tagttagcgg accgacgcgt acgcggccgc tcgagatgga
     5041 gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa
     5101 cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccgagcagg gccgcatgac
     5161 caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt
     5221 gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct
     5281 gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg
     5341 cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa
     5401 ggtgatgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag
     5461 caacgccacc gtggagcacc tgcaccccat gggcgataac gatctggatg gcagcttcac
     5521 ccgcaccttc agcctgcgcg acggcggcta ctacagctcc gtggtggaca gccacatgca
     5581 cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg
     5641 ccgcgtggag gaggatcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt
     5701 caagaccccg gatgcagatg ccggtgaaga aagagtttaa acggccggcc gcggtctgta
     5761 caagtaggat tcgtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     5821 catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat cgataatcaa
     5881 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5941 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6001 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6061 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6121 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6181 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6241 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6301 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6361 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6421 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     6481 cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     6541 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     6601 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6661 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     6721 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     6781 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     6841 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     6901 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     6961 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     7021 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7081 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7141 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc
     7201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7861 gtggaacgaa aactcacgtt aagggatttt ggtcatgatt acgccccgcc ctgccactca
     7921 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     7981 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     8041 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     8101 aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc tttagggaaa
     8161 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     8221 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     8281 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     8341 aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     8401 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     8461 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     8521 atatcaacgg tggtatatcc agtgattttt ttctccatac tcttcctttt tcaatattat
     8581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
