LOCUS       V010374                 8491 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010374
VERSION     V010374
KEYWORDS    pRFP-CB-shLenti
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8491)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8491)
  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..8491
                     /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..3272
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     CDS             3299..3694
                     /label="BSD"
                     /note="blasticidin S deaminase"
     enhancer        4104..4483
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4485..4684
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4713..4731
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4839..5531
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     protein_bind    complement(5586..5619)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5675..6263
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6266..6282)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             6792..6972
                     /label="3' LTR (truncated)"
                     /note="3' LTR (truncated)"
                     /note="truncated 3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7034..7622)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7702..8358)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(8359..8463)
                     /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 gggatctgat caagagacac gtacgaccat
     3301 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3361 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     3421 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3481 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     3541 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3601 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3661 tgaattgctg ccctctggtt atgtgtggga gggctaactg cagcgggact ctggggttcg
     3721 aaatgaccga atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     3781 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3841 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg
     3901 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3961 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4021 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaac
     4081 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
     4141 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4201 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4261 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4321 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     4381 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     4441 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     4501 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4561 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4621 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4681 cagagctcgt ttagtgaacc gtcagaattt tgtaatacga ctcactatag ggcggccggg
     4741 aattgatccg gtaccgagga gactgccgcc gcgatcgccg gcgcgccaga tctcaagctt
     4801 aactagttag cggaccgacg cgtacgcggc cgctcgagat gagcgagctg atcaaggaga
     4861 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
     4921 ccgagggcga aggcaagccc tacgagggca cccagaccat gaagatcaag gtggtcgagg
     4981 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaag
     5041 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     5101 tcacatggga gagaatcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     5161 ccagcttcca gaacggctgc atcatctaca acgtcaagat caacggggtg aacttcccat
     5221 ccaacggccc tgtgatgcag aagaaaacac gcggctggga ggccaacacc gagatgctgt
     5281 accccgctga cggcggcctg agaggccaca gccagatggc cctgaagctc gtgggcgggg
     5341 gctacctgca ctgctccttc aagaccacat acagatccaa gaaacccgct aagaacctca
     5401 agatgcccgg cttccacttc gtggaccaca gactggaaag aatcaaggag gccgacaaag
     5461 agacctacgt cgagcagcac gagatggctg tggccaagta ctgcgacctc cctagcaaac
     5521 tggggcacag atgagtttaa acggccggcc gcggtctgta caagtaggat tcgtcgaggg
     5581 acctaataac ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt
     5641 ccactaggta caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     5701 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5761 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5821 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5881 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5941 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6001 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6061 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6121 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6181 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6241 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac
     6301 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
     6361 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
     6421 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     6481 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
     6541 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
     6601 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
     6661 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
     6721 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
     6781 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6841 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6901 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6961 aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7681 aagggatttt ggtcatgatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     7741 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     7801 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     7861 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     7921 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     7981 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     8041 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     8101 atcccatatc accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat
     8161 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     8221 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     8281 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     8341 agtgattttt ttctccatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8401 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8461 gcgcacattt ccccgaaaag tgccacctga c
//
