LOCUS       V010361                 8104 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010361
VERSION     V010361
KEYWORDS    pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8104)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8104)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8104
                     /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    2195..2312
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2361..2760
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2770..3390
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     CDS             3397..3450
                     /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             3457..4167
                     /label="TagRFP"
                     /note="monomeric derivative of red fluorescent protein from
                     Entacmaea quadricolor (Merzlyak et al., 2007)"
     misc_feature    4183..4771
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4845..5078
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5150..5284
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5311..5446
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5479..5495)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5503..5519)
                     /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(5527..5557)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5572..5593)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5881..6469)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6643..7500)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7501..7605)
                     /label="AmpR promoter"
     primer_bind     8079..8095
                     /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 cttcgtgcgt
     2101 ttggggtttc ggactgtaga actctgaacc tctcggtggt cgccgtatca ttagaattct
     2161 cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2221 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2281 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2341 cactttggcc gccgttcgaa ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2401 cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc
     2461 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2521 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2581 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2641 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2701 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     2761 ctagacacca tgtccagact ggacaagagc aaagtcatca actctgcctt ggagctcctg
     2821 aatgaagttg gcattgaggg cttgaccacc aggaagctgg cccagaagct gggtgtggag
     2881 cagcctaccc tgtactggca tgtgaagaac aagagggctc tgcttgatgc cctggccatt
     2941 gagatgttgg acaggcacca cacccacttc tgccctctgg aaggggagtc ctggcaggac
     3001 ttcctgagga acaatgccaa gagcttcaga tgtgccttgc tctcccaccg ggacggtgcc
     3061 aaagttcact tgggcaccag gcctacagag aagcagtatg agaccctgga gaaccagctg
     3121 gcattcctgt gccaacaagg cttctccctg gaaaatgcct tgtatgccct ctctgctgtg
     3181 ggccacttca ccttgggctg tgtgctggag gaccaggagc accaagttgc caaggaggag
     3241 agggagaccc ccaccactga ctccatgcca ccactgctgc ggcaagctat tgagttgttt
     3301 gaccaccaag gggctgagcc tgcattcctg tttggcctgg aactgatcat ctgtggcctg
     3361 gaaaagcagc tgaaatgtga gtctggctct ggatccgagg gcagaggaag ccttctaaca
     3421 tgcggtgacg tggaggagaa tcccggccct tccggaatgg tgtctaaggg cgaagagctg
     3481 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
     3541 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     3601 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     3661 ggcagcagaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     3721 cctgagggct tcacatggga gagagtcacc acatacgagg acgggggcgt gctgaccgct
     3781 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     3841 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccaacacc
     3901 gagatgctgt accccgctga cggcggcctg gaaggcagaa gcgacatggc cctgaagctc
     3961 gtgggcgggg gccacctgat ctgcaacttc aagaccacat acagatccaa gaaacccgct
     4021 aagaacctca agatgcccgg cgtctactat gtggaccaca gactggaaag aatcaaggag
     4081 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     4141 cctagcaaac tggggcacaa acttaattga atctaggtcg acaatcaacc tctggattac
     4201 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4261 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4321 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4381 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4441 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4501 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4561 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     4621 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4681 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg
     4741 agtcggatct ctctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa
     4801 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     4861 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     4921 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     4981 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5041 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     5101 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     5161 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5221 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5281 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5341 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5401 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga
     5461 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5521 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5581 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6061 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6301 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6481 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6541 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7621 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7681 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     7741 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     7801 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     7861 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     7921 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     7981 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8041 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     8101 gctg
//
