LOCUS       V010368                 8047 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010368
VERSION     V010368
KEYWORDS    pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8047)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8047)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8047
                     /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="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"
     gap             2095..2112
                     /estimated_length=18
     misc_feature    2095..2112
                     /label="clonal barcode"
                     /note="clonal barcode"
     misc_feature    2144..2259
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     promoter        2325..2724
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2737..3447
                     /label="TagRFP"
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
     CDS             3454..3507
                     /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             3514..4110
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4126..4714
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4788..5021
                     /label="3' LTR (Delta-U3)"
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5093..5227
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5254..5389
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5422..5438)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5446..5462)
                     /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(5470..5500)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5515..5536)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5824..6412)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6586..7443)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7444..7548)
                     /label="AmpR promoter"
     primer_bind     8022..8038
                     /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 ttcgnnnnnn
     2101 nnnnnnnnnn nnttcggact gtagaactct gaacctcgag caatttaaaa gaaaaggggg
     2161 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2221 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttc tgcgttgttg tcggtgctcg
     2281 ttctctgctc ttcacgctac tgaattcatc accggttctt cgaaggcctc cgcgccgggt
     2341 tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg
     2401 gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2461 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2521 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2581 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2641 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2701 ggatcgctgt gatcgtcact tggtctagac gccaccatgg tgtctaaggg cgaagagctg
     2761 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
     2821 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     2881 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     2941 ggcagcagaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     3001 cctgagggct tcacatggga gagagtcacc acatacgagg acgggggcgt gctgaccgct
     3061 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     3121 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccaacacc
     3181 gagatgctgt accccgctga cggcggcctg gaaggcagaa gcgacatggc cctgaagctc
     3241 gtgggcgggg gccacctgat ctgcaacttc aagaccacat acagatccaa gaaacccgct
     3301 aagaacctca agatgcccgg cgtctactat gtggaccaca gactggaaag aatcaaggag
     3361 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     3421 cctagcaaac tggggcacaa acttaatgga tccgagggca gaggaagcct tctaacatgc
     3481 ggtgacgtgg aggagaatcc cggcccttcc gggatgaccg agtacaagcc cacggtgcgc
     3541 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3601 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3661 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3721 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3781 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3841 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     3901 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     3961 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4021 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4081 acctggtgca tgacccgcaa gcccggtgcc tgaatctagg tcgacaatca acctctggat
     4141 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4201 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4261 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4321 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4381 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4441 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4501 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     4561 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4621 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct tcgccctcag
     4681 acgagtcgga tctctctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     4741 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     4801 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca
     4861 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     4921 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     4981 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     5041 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     5101 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5161 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5221 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5281 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5341 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     5401 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5461 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5521 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5581 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5641 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5701 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5761 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5821 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5881 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5941 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6001 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6061 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6121 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6181 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6241 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6301 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6361 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6421 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6481 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6541 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6601 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6661 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6721 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6781 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6841 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6901 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6961 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7021 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7081 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7141 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7201 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7261 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7321 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7381 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7441 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7501 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7561 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7621 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7801 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7861 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7921 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7981 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     8041 caagctg
//
