LOCUS       pRetroX-TetOne.        8508 bp DNA     circular SYN 01-JAN-1980
DEFINITION  All-in-one retroviral vector for strong inducible expression of 
            genes using the Tet-On(R) system.
ACCESSION   .
VERSION     .
KEYWORDS    pRetroX-TetOne
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8508)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        164..543
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    974..1331
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1405..1812
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1822..2193
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     polyA_signal    2298..2432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2607..2638
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        complement(2639..3006)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     promoter        3023..3533
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3552..4295
                     /label=Tet-On(R) 3G
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
     misc_feature    4316..4719
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4880..5092
                     /label=3'-LTR del in U3
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     polyA_signal    5735..5856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5879..5895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6370..6474
                     /label=AmpR promoter
     CDS             6475..7332
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7506..8094
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8382..8403
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8418..8448
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8456..8472
                     /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)."
     primer_bind     8480..8496
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctagctt tgctcttagg agtttcctaa tacatcccaa actcaaatat ataaagcatt
       61 tgacttgttc tatgcgccta ggagcttggc ccattgcata cgttgtatcc atatcataat
      121 atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg attattgact
      181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      361 tgggtggagt atttacgcta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcat ctagagcgct
      721 tctgctcccc gagctctata aaagagccca caacccctca ctcggggcgc cagtcctccg
      781 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
      841 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
      901 ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc
      961 accaccggga ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta
     1021 tgactgattt tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga
     1081 cccgtggtgg aactgacgag ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg
     1141 acttcggggg ccgtttttgt ggcccgacct gagtcctaaa atcccgatcg tttaggactc
     1201 tttggtgcac cccccttaga ggagggatat gtggttctgg taggagacga gaacctaaaa
     1261 cagttcccgc ctccgtctga atttttgctt tcggtttggg accgaagccg cgccgcgcgt
     1321 cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     1381 gaaaatgagg gcccgggcta gcctgttacc actcccttaa gtttgacctt aggtcactgg
     1441 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     1501 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     1561 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     1621 ccagaccagg tggggtacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     1681 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     1741 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     1801 tctctaggcg cccccatatg gccatatgag atcctatatg gggcaccccc gccccttgta
     1861 aacttccctg accctgacat gacaagagtt actaacagcc cctctctcca agctcactta
     1921 caggctctct acttagtcca gcacgaagtc tggagacctc tggcggcagc ctaccaagaa
     1981 caactggacc gaccggtggt acctcaccct taccgagtcg gcgacacagt gtgggtccgc
     2041 cgacaccaga ctaagaacct agaacctcgc tggaaaggac cttacacagt cctgctgacc
     2101 acccccaccg ccctcaaagt agacggcatc gcagcttgga tacacgccgc ccacgtgaag
     2161 gctgccgacc ccgggggtgg accatcctct agatcataat cagccatacc acatttgtag
     2221 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2281 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaggcaata
     2341 gcatcacaaa tttcacaaat aaggcatttt tttcactgca ttctagtttt ggtttgtcca
     2401 aactcatcaa tgtatcttat catgtctgga tctcaaatcc ctcggaagct gcgcctgtct
     2461 taggttggag tgatacattt ttatcacttt tacccgtctt tggattaggc agtagctctg
     2521 acggccctcc tgtcttaggt tagtgaaaaa tgtcactctc ttacccgtca ttggctgtcc
     2581 agcttagctc gcaggggagg tggtctggat ccgccggcac cggtgtatac gggaattctt
     2641 tacgagggta ggaagtggta cggaaagttg gtataagaca aaagtgttgt ggaattgaag
     2701 tttactcaaa aaatcagcac tcttttatag gcgccctggt ttacataagc aaagcttata
     2761 cgttctctat cactgatagg gagtaaactg gatatacgtt ctctatcact gatagggagt
     2821 aaactgtaga tacgttctct atcactgata gggagtaaac tggtcatacg ttctctatca
     2881 ctgataggga gtaaactcct tatacgttct ctatcactga tagggagtaa agtctgcata
     2941 cgttctctat cactgatagg gagtaaactc ttcatacgtt ctctatcact gatagggagt
     3001 aaactcgagg tgataattcc acggggttgg ggttgcgcct tttccaaggc agccctgggt
     3061 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     3121 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     3181 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     3241 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     3301 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     3361 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     3421 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     3481 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     3541 atcgaattac catgtctaga ctggacaaga gcaaagtcat aaactctgct ctggaattac
     3601 tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     3661 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     3721 tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag tcatggcaag
     3781 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     3841 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     3901 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     3961 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     4021 aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca attgagctgt
     4081 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     4141 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     4201 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     4261 ctcttgacga ttttgacctt gacatgctcc ccgggtaact aagtaagatc ctgttaatca
     4321 acctctggat tacaaaattt gtgaaagatt gactgatatt cttaactatg ttgctccttt
     4381 tacgctgtgt ggatatgctg ctttaatgcc tctgtatcat gctattgctt cccgtacggc
     4441 tttcgttttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4501 cgttgtccgt caacgtggcg tggtgtgctc tgtgtttgct gacgcaaccc ccactggctg
     4561 gggcattgcc accacctgtc aactcctttc tgggactttc gctttccccc tcccgatcgc
     4621 cacggcagaa ctcatcgccg cctgccttgc ccgctgctgg acaggggcta ggttgctggg
     4681 cactgataat tccgtggtgt tgtcggggaa gctgacgtcg attcctgcgg agatatcagt
     4741 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
     4801 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4861 ctgtaggttt ggcaagctag cttctcgctt ctgttcgcgc gcttctgctc ccagacatca
     4921 acaaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     4981 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5041 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacgcgtatg
     5101 ccgcatagtt aagccaccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5161 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5221 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5281 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5341 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5401 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5461 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     5521 ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct ccgaccgaag ccgacccggg
     5581 cggccccgcc gaccccgcac ccgcccccga ggcccaccga ctctagagga tcataatcag
     5641 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5701 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5761 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5821 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat ggcgcgtcga attaattcac
     5881 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5941 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6001 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     6061 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     6121 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6181 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6241 agaggttttc accgtcatca ccgaaacgcg cgatgacgaa agggcctcgt gatacgccta
     6301 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6361 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6421 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6481 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6541 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6601 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6661 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6721 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6781 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6841 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6901 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6961 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7021 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7081 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7141 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7201 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7261 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7321 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7381 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7441 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7501 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7561 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7621 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     7681 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7741 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7801 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7861 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     7921 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7981 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8041 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8101 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8161 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8221 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8281 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8341 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8401 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8461 agcggataac aatttcacac aggaaacagc tatgaccatg attacgcc
//
