LOCUS       Cellecta-pRSITER        8198 bp ds-DNA     circular SYN 02-OCT-2017
DEFINITION  Lentiviral vector for inducible shRNA expression, with NO NGS 
            cassette, plus TagRFP fluorescent protein.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8198)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2017 from SnapGene 4.0.5 to Vector NTI(R) format
            http://www.snapgene.com
COMMENT     NO NGS cassette
            NOT Compatible with shRNA Library Construction
COMMENT     VNTDATE|309830400|
COMMENT     VNTDBDATE|818553600|
COMMENT     VNTNAME|Cellecta-pRSITER-U6Tet-sh-EF1-TetRep-2A-TagRFP.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
     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"
     promoter        2345..2858
                     /label=EF1
                     /note="/vntifkey=29"
     CDS             2870..3490
                     /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             3497..3550
                     /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             3557..4270
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
     misc_feature    4277..4865
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4939..5172
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5244..5365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5405..5540
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5975..6563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6734..7594)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(7595..7699)
                     /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 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 tccggaatgg tgtctaaggg cgaagagctg attaaggaga acatgcacat
     3601 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     3661 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     3721 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagaa ccttcatcaa
     3781 ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga
     3841 gagagtcacc acatacgagg acgggggcgt gctgaccgct acccaggaca ccagcctcca
     3901 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
     3961 tgtgatgcag aagaaaacac tcggctggga ggccaacacc gagatgctgt accccgctga
     4021 cggcggcctg gaaggcagaa gcgacatggc cctgaagctc gtgggcgggg gccacctgat
     4081 ctgcaacttc aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
     4141 cgtctactat gtggaccaca gactggaaag aatcaaggag gccgacaaag agacctacgt
     4201 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacaa
     4261 acttaattag gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4321 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4381 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4441 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4501 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4561 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4621 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4681 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4741 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4801 gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctctcttt gggccgcctc
     4861 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     4921 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa gatctgcttt
     4981 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5041 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     5101 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     5161 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     5221 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     5281 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5341 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     5401 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5461 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5521 aggaggcttt tttggaggcc tagacttttg cagagaccaa attcgtaatc atgtcatagc
     5581 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5641 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5701 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5761 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6361 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6481 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6601 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6661 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6721 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6781 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6841 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6901 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6961 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7021 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7081 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7141 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7201 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7261 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7321 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7381 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7441 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7501 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7561 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7621 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7681 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7741 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7801 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7861 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7921 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7981 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     8041 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     8101 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     8161 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
