LOCUS       Cellecta-pRSITPR        8887 bp ds-DNA     circular SYN 13-SEP-2019
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8887)
  AUTHORS   Cellecta
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 13, 2019 from SnapGene 4.3.11 to Vector NTI(R) 
            format
            https://www.snapgene.com
COMMENT     VNTDATE|712368000|
COMMENT     VNTDBDATE|888624000|
COMMENT     VNTNAME|Cellecta-pRSITPRP-U6Tet-sh-HTS3-PGK-TetRep-2A-TagRFP-2A-Puro
            |
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=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    2195..2312
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2365..2865
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2878..3498
                     /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             3505..3558
                     /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             3565..4275
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
     CDS             4294..4347
                     /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             4354..4953
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     misc_feature    4966..5554
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5628..5861
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5933..6054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6094..6229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(6664..7252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7423..8283)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(8284..8388)
                     /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 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 ggggttgcgc cttttccaag gcagccctgg gtttgcgcag
     2401 ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc
     2461 acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc
     2521 ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc
     2581 cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag
     2641 ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg
     2701 cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg
     2761 tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg
     2821 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagggtct agacaccatg
     2881 tccagactgg acaagagcaa agtcatcaac tctgccttgg agctcctgaa tgaagttggc
     2941 attgagggct tgaccaccag gaagctggcc cagaagctgg gtgtggagca gcctaccctg
     3001 tactggcatg tgaagaacaa gagggctctg cttgatgccc tggccattga gatgttggac
     3061 aggcaccaca cccacttctg ccctctggaa ggggagtcct ggcaggactt cctgaggaac
     3121 aatgccaaga gcttcagatg tgccttgctc tcccaccggg acggtgccaa agttcacttg
     3181 ggcaccaggc ctacagagaa gcagtatgag accctggaga accagctggc attcctgtgc
     3241 caacaaggct tctccctgga aaatgccttg tatgccctct ctgctgtggg ccacttcacc
     3301 ttgggctgtg tgctggagga ccaggagcac caagttgcca aggaggagag ggagaccccc
     3361 accactgact ccatgccacc actgctgcgg caagctattg agttgtttga ccaccaaggg
     3421 gctgagcctg cattcctgtt tggcctggaa ctgatcatct gtggcctgga aaagcagctg
     3481 aaatgtgagt ctggctcttt cgaagagggc agaggaagcc ttctaacatg cggtgacgtg
     3541 gaggagaatc ccggcccttc cggaatggtg tctaagggcg aagagctgat taaggagaac
     3601 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
     3661 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     3721 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc
     3781 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc
     3841 acatgggaga gagtcaccac atacgaggac gggggcgtgc tgaccgctac ccaggacacc
     3901 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
     3961 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga gatgctgtac
     4021 cccgctgacg gcggcctgga aggcagaagc gacatggccc tgaagctcgt gggcgggggc
     4081 cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag
     4141 atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag
     4201 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
     4261 gggcacaaac ttaatgctag caccggtaga tccgagggca gaggaagcct tctaacatgc
     4321 ggtgacgtgg aggagaatcc cggcccttcc gggatgaccg agtacaagcc cacggtgcgc
     4381 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4441 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4501 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4561 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4621 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4681 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4741 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4801 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4861 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4921 acctggtgca tgacccgcaa gcccggtgcc tgaatctagg tcgacaatca acctctggat
     4981 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5041 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5101 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5161 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5221 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5281 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5341 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5401 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5461 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct tcgccctcag
     5521 acgagtcgga tctctctttg ggccgcctcc ccgcctggta cctttaagac caatgactta
     5581 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5641 tcactcccaa cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca
     5701 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     5761 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     5821 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     5881 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     5941 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6001 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6061 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6121 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6181 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6241 agagaccaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8401 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8461 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8521 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8581 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     8641 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     8701 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8761 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     8821 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     8881 caagctg
//
