LOCUS       pRetroX-TetOne-P        9463 bp DNA     circular SYN 01-JAN-1980
DEFINITION  All-in-one retroviral vector with a puromycin resistance marker, for
            strong inducible expression of genes using the Tet-On(R) system.
ACCESSION   .
VERSION     .
KEYWORDS    pRetroX-TetOne-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9463)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9463)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9463
                     /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"
     promoter        4316..4645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4654..5250
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    5271..5674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5835..6047
                     /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    6690..6811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6834..6850)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7325..7429
                     /label=AmpR promoter
     CDS             7430..8287
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8461..9049
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9337..9358
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9373..9403
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9411..9427
                     /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     9435..9451
                     /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 aagtaagaat tgaatgtgtg
     4321 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4381 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4441 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4501 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4561 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4621 ttttggaggc ctaggctttt gcaaaacgcg accatgaccg agtacaagcc cacggtgcgc
     4681 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     4741 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     4801 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4861 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4921 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4981 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5041 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5101 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5161 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5221 acctggtgca tgacccgcaa gcccggtgcc tgaacgcgta attccctgtt aatcaacctc
     5281 tggattacaa aatttgtgaa agattgactg atattcttaa ctatgttgct ccttttacgc
     5341 tgtgtggata tgctgcttta atgcctctgt atcatgctat tgcttcccgt acggctttcg
     5401 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5461 tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc aacccccact ggctggggca
     5521 ttgccaccac ctgtcaactc ctttctggga ctttcgcttt ccccctcccg atcgccacgg
     5581 cagaactcat cgccgcctgc cttgcccgct gctggacagg ggctaggttg ctgggcactg
     5641 ataattccgt ggtgttgtcg gggaagctga cgtcgattcc tgcggagata tcagtggtcc
     5701 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     5761 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     5821 ggtttggcaa gctagcttct cgcttctgtt cgcgcgcttc tgctcccaga catcaacaaa
     5881 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     5941 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     6001 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacgc gtatgccgca
     6061 tagttaagcc accgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     6121 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     6181 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6241 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     6301 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     6361 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6421 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     6481 gcgcccgacc gaaaggagcg cacgacccca tggctccgac cgaagccgac ccgggcggcc
     6541 ccgccgaccc cgcacccgcc cccgaggccc accgactcta gaggatcata atcagccata
     6601 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     6661 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     6721 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6781 gtggtttgtc caaactcatc aatgtatctt atcatggcgc gtcgaattaa ttcactggcc
     6841 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6901 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6961 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     7021 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     7081 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     7141 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     7201 ttttcaccgt catcaccgaa acgcgcgatg acgaaagggc ctcgtgatac gcctattttt
     7261 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7321 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7381 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7441 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7501 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7561 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7621 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7681 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7741 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7801 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     7861 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     7921 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     7981 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8041 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8101 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     8161 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8221 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8281 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8341 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8581 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     8641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8821 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     8881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     9061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     9121 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     9181 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     9241 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     9301 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     9361 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     9421 ataacaattt cacacaggaa acagctatga ccatgattac gcc
//
