LOCUS       V010362                 8090 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010362
VERSION     V010362
KEYWORDS    pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8090)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8090)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            Not suitable for high-throughput sequencing.
FEATURES             Location/Qualifiers
     source          1..8090
                     /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"
     protein_bind    2020..2037
                     /gene="tetO"
                     /label="tetracycline repressor TetR binding site"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2046..2064
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2066..2095
                     /label="cloning site"
                     /note="cloning site"
                     /note="shRNA cloning site"
     misc_feature    2179..2296
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2345..2556
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2569..2837
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2870..3490
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     CDS             3497..3550
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3557..4153
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4169..4757
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4831..5064
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5136..5270
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5297..5432
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5465..5481)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5489..5505)
                     /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(5513..5543)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5558..5579)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5867..6455)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6629..7486)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7487..7591)
                     /label="AmpR promoter"
     primer_bind     8065..8081
                     /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 acttgaaagc cctatcagtg atagagattt
     2041 atatatccct atcagtgata gagacaccgg agtcttcttt tttgaagaca cttcggactg
     2101 tagaactctg aacctctcgg tggtcgccgt atcattagaa ttctcgacct cgagacaaat
     2161 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2221 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2281 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggccgccgac
     2341 tagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     2401 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     2461 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     2521 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     2581 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     2641 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     2701 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     2761 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     2821 ctgcgccgtt acagatccaa gctgtgaccg gcgcctactt ctagacacca tgtccagact
     2881 ggacaagagc aaagtcatca actctgcctt ggagctcctg aatgaagttg gcattgaggg
     2941 cttgaccacc aggaagctgg cccagaagct gggtgtggag cagcctaccc tgtactggca
     3001 tgtgaagaac aagagggctc tgcttgatgc cctggccatt gagatgttgg acaggcacca
     3061 cacccacttc tgccctctgg aaggggagtc ctggcaggac ttcctgagga acaatgccaa
     3121 gagcttcaga tgtgccttgc tctcccaccg ggacggtgcc aaagttcact tgggcaccag
     3181 gcctacagag aagcagtatg agaccctgga gaaccagctg gcattcctgt gccaacaagg
     3241 cttctccctg gaaaatgcct tgtatgccct ctctgctgtg ggccacttca ccttgggctg
     3301 tgtgctggag gaccaggagc accaagttgc caaggaggag agggagaccc ccaccactga
     3361 ctccatgcca ccactgctgc ggcaagctat tgagttgttt gaccaccaag gggctgagcc
     3421 tgcattcctg tttggcctgg aactgatcat ctgtggcctg gaaaagcagc tgaaatgtga
     3481 gtctggctct ggatccgagg gcagaggaag ccttctaaca tgcggtgacg tggaggagaa
     3541 tcccggccct tccgggatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3601 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3661 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3721 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3781 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3841 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3901 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3961 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4021 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4081 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4141 caagcccggt gcctgaatct aggtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     4201 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4261 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4321 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4381 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4441 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4501 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4561 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4621 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4681 gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctctct
     4741 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     4801 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat
     4861 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     4921 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     4981 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5041 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     5101 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     5161 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5221 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     5281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5401 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacc aaattcgtaa
     5461 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6301 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7621 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7681 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     7741 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7801 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7861 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7921 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7981 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     8041 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
