LOCUS       V010370                 8058 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010370
VERSION     V010370
KEYWORDS    pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8058)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8058)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8058
                     /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    2138..2255
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2336..2735
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2748..3458
                     /label="TagRFP"
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
     CDS             3465..3518
                     /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             3525..4121
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4137..4725
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4799..5032
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5104..5238
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5265..5400
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5433..5449)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5457..5473)
                     /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(5481..5511)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5526..5547)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5835..6423)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6597..7454)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7455..7559)
                     /label="AmpR promoter"
     primer_bind     8033..8049
                     /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 acctctgaat tctcactcga gccacaattt taaaagaaaa ggggggattg
     2161 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2221 aattacaaaa acaaattaca aaaattcaaa attttgcggt tgtgcttttt gcgtgtaaat
     2281 ctcggtggtc gccgtatcat ggcttgtgcg atgcgttggc taccggttct tcgaaggcct
     2341 ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag cgctgccacg
     2401 tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2461 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2521 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2581 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2641 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2701 ttcttgtttg tggatcgctg tgatcgtcac ttggtctaga cgccaccatg gtgtctaagg
     2761 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     2821 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2881 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     2941 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta
     3001 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgag gacgggggcg
     3061 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     3121 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     3181 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga agcgacatgg
     3241 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     3301 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     3361 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     3421 actgcgacct ccctagcaaa ctggggcaca aacttaatgg atccgagggc agaggaagcc
     3481 ttctaacatg cggtgacgtg gaggagaatc ccggcccttc cgggatgacc gagtacaagc
     3541 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     3601 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     3661 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     3721 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     3781 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     3841 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     3901 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     3961 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4021 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4081 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaatctag gtcgacaatc
     4141 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4201 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4261 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4321 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4381 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4441 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4501 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4561 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4621 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtctccgcc
     4681 ttcgccctca gacgagtcgg atctctcttt gggccgcctc cccgcctggt acctttaaga
     4741 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4801 gaagggctaa ttcactccca acgaagataa gatctgcttt ttgcttgtac tgggtctctc
     4861 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4921 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4981 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     5041 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     5101 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5221 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     5281 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5341 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5401 tagacttttg cagagaccaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     5461 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5521 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5581 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5641 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5701 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5821 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5881 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5941 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6001 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6061 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6121 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6181 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6241 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6301 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6361 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6421 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6481 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6541 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6601 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6661 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6721 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6781 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6841 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6901 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6961 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7021 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7081 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7141 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7201 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7261 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7321 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7381 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7441 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7501 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7561 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7621 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     7681 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     7741 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     7801 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     7861 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     7921 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7981 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8041 acggccagtg ccaagctg
//
