LOCUS       V010367                 8028 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010367
VERSION     V010367
KEYWORDS    pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8028)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8028)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label="5' LTR (truncated)"
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1078..1311
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1496..1540
                     /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             1689..1730
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        1819..2059
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2065..2094
                     /label="cloning site"
                     /note="cloning site"
                     /note="shRNA cloning site"
     misc_feature    2122..2237
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     promoter        2303..2702
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2715..3428
                     /label="TagGFP2"
                     /note="monomeric green fluorescent protein, also known as
                     mTagGFP"
     CDS             3435..3488
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label="2A peptide from Thosea asigna virus capsid protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3495..4091
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4107..4695
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4769..5002
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5074..5208
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5235..5370
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5403..5419)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5427..5443)
                     /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)."
     promoter        complement(5451..5481)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5496..5517)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5805..6393)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6567..7424)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7425..7529)
                     /label="AmpR promoter"
     primer_bind     8003..8019
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 atggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga
     1861 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     1921 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     1981 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     2041 tatatcttgt ggaaaggacg aaacaccgga gtcttctttt ttgaagacac ttcggactgt
     2101 agaactctga acctcgagca atttaaaaga aaagggggga ttggggggta cagtgcaggg
     2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2221 acaaaaattc aaaatttctg cgttgttgtc ggtgctcgtt ctctgctctt cacgctactg
     2281 aattcatcac cggttcttcg aaggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     2341 cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg tcctgatcct
     2401 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     2461 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     2521 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     2581 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     2641 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     2701 gtctagacgc caccatgagc gggggcgagg agctgttcgc cggcatcgtg cccgtgctga
     2761 tcgagctgga cggcgacgtg cacggccaca agttcagcgt gcgcggcgag ggcgagggcg
     2821 acgccgacta cggcaagctg gagatcaagt tcatctgcac caccggcaag ctgcccgtgc
     2881 cctggcccac cctggtgacc accctctgct acggcatcca gtgcttcgcc cgctaccccg
     2941 agcacatgaa gatgaacgac ttcttcaaga gcgccatgcc cgagggctac atccaggagc
     3001 gcaccatcca gttccaggac gacggcaagt acaagacccg cggcgaggtg aagttcgagg
     3061 gcgacaccct ggtgaaccgc atcgagctga agggcaagga cttcaaggag gacggcaaca
     3121 tcctgggcca caagctggag tacagcttca acagccacaa cgtgtacatc cgccccgaca
     3181 aggccaacaa cggcctggag gctaacttca agacccgcca caacatcgag ggcggcggcg
     3241 tgcagctggc cgaccactac cagaccaacg tgcccctggg cgacggcccc gtgctgatcc
     3301 ccatcaacca ctacctgagc actcagacca agatcagcaa ggaccgcaac gaggcccgcg
     3361 accacatggt gctcctggag tccttcagcg cctgctgcca cacccacggc atggacgagc
     3421 tgtacagggg atccgagggc agaggaagcc ttctaacatg cggtgacgtg gaggagaatc
     3481 ccggcccttc cgggatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     3541 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     3601 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     3661 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     3721 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     3781 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     3841 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     3901 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     3961 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     4021 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     4081 agcccggtgc ctgaatctag gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     4141 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4201 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4261 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4321 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4381 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     4441 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     4501 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     4561 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4621 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctctcttt
     4681 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     4741 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa
     4801 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     4861 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     4921 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     4981 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     5041 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5101 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5161 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     5221 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5281 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5341 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa attcgtaatc
     5401 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5461 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5521 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5581 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5641 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5701 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5761 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5821 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5881 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5941 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6001 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6061 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6241 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6361 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6421 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6481 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6541 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6781 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6841 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7081 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7201 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7441 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7501 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7561 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7621 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7681 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     7741 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7801 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7861 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7921 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7981 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
