LOCUS       Cellecta-pRSITEP        8120 bp ds-DNA     circular SYN 05-JAN-2018
DEFINITION  Lentiviral vector for inducible shRNA expression, with NO NGS 
            cassette, plus puromycin resistance marker.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8120)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2018 from SnapGene 4.1.3 to Vector NTI(R) format
            http://www.snapgene.com
COMMENT     NO NGS cassette
            NOT Compatible with shRNA Library Construction
COMMENT     VNTDATE|675302400|
COMMENT     VNTDBDATE|829872000|
COMMENT     VNTNAME|Cellecta-pRSITEP-U6Tet-sh-EF1-TetRep-2A-Puro.dna|
COMMENT     VNTAUTHORNAME|Cellecta|
FEATURES             Location/Qualifiers
     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."
     promoter        1803..2064
                     /label=U6T8T
                     /label=U6-Tet
     primer          1863..1888
                     /label=FwdU6-1
                     /note="sequence: caaggctgttagagagataattggaa"
     primer          1914..1939
                     /label=FwdU6-3
                     /note="sequence: ATTAGTACAAAATACGTGACGTAGAA"
     protein_bind    2020..2037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2046..2064
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2066..2095
                     /label=cloning site
     misc_feature    2179..2296
                     /label=cPPT
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer          complement(2228..2253)
                     /label=Rev-cPPT10
                     /note="sequence: tgtatgtctgttgctattatgtctac"
     promoter        2375..2888
                     /label=EF1
     CDS             2900..3520
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
     CDS             3527..3580
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
     CDS             3587..4186
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    4199..4787
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4861..5094
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5166..5287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5327..5462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5897..6485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6656..7516)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(7517..7621)
                     /gene="bla"
                     /label=AmpR promoter
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 tagtggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     2401 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     2461 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     2521 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     2581 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     2641 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     2701 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     2761 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     2821 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     2881 gcgcctactt ctagacacca tgtccagact ggacaagagc aaagtcatca actctgcctt
     2941 ggagctcctg aatgaagttg gcattgaggg cttgaccacc aggaagctgg cccagaagct
     3001 gggtgtggag cagcctaccc tgtactggca tgtgaagaac aagagggctc tgcttgatgc
     3061 cctggccatt gagatgttgg acaggcacca cacccacttc tgccctctgg aaggggagtc
     3121 ctggcaggac ttcctgagga acaatgccaa gagcttcaga tgtgccttgc tctcccaccg
     3181 ggacggtgcc aaagttcact tgggcaccag gcctacagag aagcagtatg agaccctgga
     3241 gaaccagctg gcattcctgt gccaacaagg cttctccctg gaaaatgcct tgtatgccct
     3301 ctctgctgtg ggccacttca ccttgggctg tgtgctggag gaccaggagc accaagttgc
     3361 caaggaggag agggagaccc ccaccactga ctccatgcca ccactgctgc ggcaagctat
     3421 tgagttgttt gaccaccaag gggctgagcc tgcattcctg tttggcctgg aactgatcat
     3481 ctgtggcctg gaaaagcagc tgaaatgtga gtctggctct ggatccgagg gcagaggaag
     3541 ccttctaaca tgcggtgacg tggaggagaa tcccggccct tccgggatga ccgagtacaa
     3601 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3661 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3721 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3781 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3841 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3901 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3961 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4021 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     4081 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     4141 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaatct aggtcgacaa
     4201 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4261 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4321 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4381 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4441 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4501 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4561 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     4621 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4681 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtctccg
     4741 ccttcgccct cagacgagtc ggatctctct ttgggccgcc tccccgcctg gtacctttaa
     4801 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     4861 tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc
     4921 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4981 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5041 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     5101 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     5161 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5221 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5281 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     5341 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5401 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5461 cctagacttt tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5641 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5701 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5761 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5821 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5881 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5941 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6001 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6061 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6121 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6181 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6241 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6301 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6361 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6421 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7081 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7621 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     7681 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     7741 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7801 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     7861 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     7921 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     7981 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     8041 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     8101 cgacggccag tgccaagctg
//
